Solid forms of 2-(4-chlorophenyl)-N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-2,2-difluoroacetamide, and their pharmaceutical compositions and uses

ABSTRACT

Solid forms comprising 2-(4-chlorophenyl)-N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-2,2-difluoroacetamide, compositions comprising the solid forms, methods of making the solid forms and methods of their uses are disclosed.

1. CROSS REFERENCE TO RELATED APPLICATIONS

This application is a divisional of U.S. application Ser. No. 15/400,630filed Jan. 6, 2017, currently allowed, which claims the benefit of thepriority of U.S. Provisional Application No. 62/276,750, filed Jan. 8,2016, the disclosures of each of which are incorporated herein byreference in their entireties.

2. FIELD

Provided herein are solid forms of2-(4-chlorophenyl)-N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-2,2-difluoroacetamide,pharmaceutical compositions thereof, and methods of their uses for thetreatment of diseases or disorders.

3. BACKGROUND OF THE DISCLOSURE 3.1 Pathobiology of Cancer and OtherDiseases

Cancer is characterized primarily by an increase in the number ofabnormal cells derived from a given normal tissue, invasion of adjacenttissues by these abnormal cells, or lymphatic or blood-borne spread ofmalignant cells to regional lymph nodes and to distant sites(metastasis). Clinical data and molecular biologic studies indicate thatcancer is a multistep process that begins with minor preneoplasticchanges, which may under certain conditions progress to neoplasia. Theneoplastic lesion may evolve clonally and develop an increasing capacityfor invasion, growth, metastasis, and heterogeneity, especially underconditions in which the neoplastic cells escape the host's immunesurveillance. Roitt, I., Brostoff, J and Kale, D., Immunology,17.1-17.12 (3rd ed., Mosby, St. Louis, Mo., 1993).

There is an enormous variety of cancers which are described in detail inthe medical literature. Examples include cancers of the lung, colon,rectum, prostate, breast, brain, and intestine. The incidence of cancercontinues to climb as the general population ages, as new cancersdevelop, and as susceptible populations (e.g., people infected with AIDSor excessively exposed to sunlight) grow. However, options for thetreatment of cancer are limited. For example, in the case of bloodcancers (e.g., multiple myeloma), few treatment options are available,especially when conventional chemotherapy fails and bone-marrowtransplantation is not an option. A tremendous demand therefore existsfor new methods and compositions that can be used to treat patients withcancer.

Many types of cancers are associated with new blood vessel formation, aprocess known as angiogenesis. Several of the mechanisms involved intumor-induced angiogenesis have been elucidated. The most direct ofthese mechanisms is the secretion by the tumor cells of cytokines withangiogenic properties. Examples of these cytokines include acidic andbasic fibroblastic growth factor (a,b-FGF), angiogenin, vascularendothelial growth factor (VEGF), and TNF-α. Alternatively, tumor cellscan release angiogenic peptides through the production of proteases andthe subsequent breakdown of the extracellular matrix where somecytokines are stored (e.g., b-FGF). Angiogenesis can also be inducedindirectly through the recruitment of inflammatory cells (particularlymacrophages) and their subsequent release of angiogenic cytokines (e.g.,TNF-α, b-FGF).

A variety of other diseases and disorders are also associated with, orcharacterized by, undesired angiogenesis. For example, enhanced orunregulated angiogenesis has been implicated in a number of diseases andmedical conditions including, but not limited to, ocular neovasculardiseases, choroidal neovascular diseases, retina neovascular diseases,rubeosis (neovascularization of the angle), viral diseases, geneticdiseases, inflammatory diseases, allergic diseases, and autoimmunediseases. Examples of such diseases and conditions include, but are notlimited to, diabetic retinopathy, retinopathy of prematurity, cornealgraft rejection, neovascular glaucoma, retrolental fibroplasia,arthritis, and proliferative vitreoretinopathy.

Accordingly, compounds that can control angiogenesis or inhibit theproduction of certain cytokines, including TNFα, may be useful in thetreatment and prevention of various diseases and conditions.

3.2 Methods of Treating Cancer

Current cancer therapy may involve surgery, chemotherapy, hormonaltherapy and/or radiation treatment to eradicate neoplastic cells in apatient (see, e.g., Stockdale, 1998, Medicine, vol. 3, Rubenstein andFederman, eds., Chapter 12, Section IV). Recently, cancer therapy couldalso involve biological therapy or immunotherapy. All of theseapproaches pose significant drawbacks for the patient. Surgery, forexample, may be contraindicated due to the health of a patient or may beunacceptable to the patient. Additionally, surgery may not completelyremove neoplastic tissue. Radiation therapy is only effective when theneoplastic tissue exhibits a higher sensitivity to radiation than normaltissue. Radiation therapy can also often elicit serious side effects.Hormonal therapy is rarely given as a single agent. Although hormonaltherapy can be effective, it is often used to prevent or delayrecurrence of cancer after other treatments have removed the majority ofcancer cells. Biological therapies and immunotherapies are limited innumber and may produce side effects such as rashes or swellings,flu-like symptoms, including fever, chills and fatigue, digestive tractproblems or allergic reactions.

With respect to chemotherapy, there is a variety of chemotherapeuticagents available for treatment of cancer. A majority of cancerchemotherapeutics act by inhibiting DNA synthesis, either directly orindirectly by inhibiting the biosynthesis of deoxyribonucleotidetriphosphate precursors, to prevent DNA replication and concomitant celldivision. Gilman et al., Goodman and Gilman's: The Pharmacological Basisof Therapeutics, Tenth Ed. (McGraw Hill, New York).

Despite availability of a variety of chemotherapeutic agents,chemotherapy has many drawbacks. Stockdale, Medicine, vol. 3, Rubensteinand Federman, eds., ch. 12, sect. 10, 1998. Almost all chemotherapeuticagents are toxic, and chemotherapy causes significant, and oftendangerous side effects including severe nausea, bone marrow depression,and immunosuppression. Additionally, even with administration ofcombinations of chemotherapeutic agents, many tumor cells are resistantor develop resistance to the chemotherapeutic agents. In fact, thosecells resistant to the particular chemotherapeutic agents used in thetreatment protocol often prove to be resistant to other drugs, even ifthose agents act by different mechanism from those of the drugs used inthe specific treatment. This phenomenon is referred to as pleiotropicdrug or multidrug resistance. Because of the drug resistance, manycancers prove or become refractory to standard chemotherapeutictreatment protocols.

Other diseases or conditions associated with, or characterized by,undesired angiogenesis are also difficult to treat. However, somecompounds such as protamine, hepain and steroids have been proposed tobe useful in the treatment of certain specific diseases. Taylor et al.,Nature 297:307 (1982); Folkman et al., Science 221:719 (1983); and U.S.Pat. Nos. 5,001,116 and 4,994,443.

Still, there is a significant need for safe and effective methods oftreating, preventing and managing cancer and other diseases andconditions, including for diseases that are refractory to standardtreatments, such as surgery, radiation therapy, chemotherapy andhormonal therapy, while reducing or avoiding the toxicities and/or sideeffects associated with the conventional therapies.

3.3 Solid Forms

The preparation and selection of a solid form of a pharmaceuticalcompound is complex, given that a change in solid form may affect avariety of physical and chemical properties, which may provide benefitsor drawbacks in processing, formulation, stability and bioavailability,among other important pharmaceutical characteristics. Potentialpharmaceutical solids include crystalline solids and amorphous solids.Amorphous solids are characterized by a lack of long-range structuralorder, whereas crystalline solids are characterized by structuralperiodicity. The desired class of pharmaceutical solid depends upon thespecific application; amorphous solids are sometimes selected on thebasis of, e.g., an enhanced dissolution profile, while crystallinesolids may be desirable for properties such as, e.g., physical orchemical stability (see, e.g., S. R. Vippagunta et al., Adv. Drug.Deliv. Rev., (2001) 48:3-26; L. Yu, Adv. Drug. Deliv. Rev., (2001)48:27-42).

Whether crystalline or amorphous, potential solid forms of apharmaceutical compound include single-component and multiple-componentsolids. Single-component solids consist essentially of thepharmaceutical compound in the absence of other compounds. Variety amongsingle-component crystalline materials may potentially arise, e.g., fromthe phenomenon of polymorphism, wherein multiple three-dimensionalarrangements exist for a particular pharmaceutical compound (see, e.g.,S. R. Byrn et al., Solid State Chemistry of Drugs, (1999) SSCI, WestLafayette). The importance of studying polymorphs was underscored by thecase of Ritonavir, an HIV protease inhibitor that was formulated as softgelatin capsules. About two years after the product was launched, theunanticipated precipitation of a new, less soluble polymorph in theformulation necessitated the withdrawal of the product from the marketuntil a more consistent formulation could be developed (see S. R.Chemburkar et al., Org. Process Res. Dev., (2000) 4:413-417).

Additional diversity among the potential solid forms of a pharmaceuticalcompound may arise, e.g., from the possibility of multiple-componentsolids. Crystalline solids comprising two or more ionic species may betermed salts (see, e.g., Handbook of Pharmaceutical Salts: Properties,Selection and Use, P. H. Stahl and C. G. Wermuth, Eds., (2002), Wiley,Weinheim). Additional types of multiple-component solids that maypotentially offer other property improvements for a pharmaceuticalcompound or salt thereof include, e.g., hydrates, solvates, co-crystalsand clathrates, among others (see, e.g., S. R. Byrn et al., Solid StateChemistry of Drugs, (1999) SSCI, West Lafayette). Moreover,multiple-component crystal forms may potentially be susceptible topolymorphism, wherein a given multiple-component composition may existin more than one three-dimensional crystalline arrangement. Thepreparation of solid forms is of great importance in the development ofa safe, effective, stable and marketable pharmaceutical compound.

Notably, it is not possible to predict a priori if crystalline forms ofa compound even exist, let alone how to successfully prepare them (see,e.g., Braga and Grepioni, 2005, “Making crystals from crystals: a greenroute to crystal engineering and polymorphism,” Chem. Commun.: 3635-3645(with respect to crystal engineering, if instructions are not veryprecise and/or if other external factors affect the process, the resultcan be unpredictable); Jones et al., 2006, Pharmaceutical Cocrystals: AnEmerging Approach to Physical Property Enhancement,” MRS Bulletin31:875-879 (At present it is not generally possible to computationallypredict the number of observable polymorphs of even the simplestmolecules); Price, 2004, “The computational prediction of pharmaceuticalcrystal structures and polymorphism,” Advanced Drug Delivery Reviews56:301-319 (“Price”); and Bernstein, 2004, “Crystal Structure Predictionand Polymorphism,” ACA Transactions 39:14-23 (a great deal still needsto be learned and done before one can state with any degree ofconfidence the ability to predict a crystal structure, much lesspolymorphic forms)).

The variety of possible solid forms creates potential diversity inphysical and chemical properties for a given pharmaceutical compound.The discovery and selection of solid forms are of great importance inthe development of an effective, stable and marketable pharmaceuticalproduct.

Citation or identification of any reference in Section 2 of thisapplication is not to be construed as an admission that the reference isprior art to the present application.

Provided herein are solid forms of2-(4-chlorophenyl)-N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-2,2-difluoroacetamide(“Compound 1”). Compound 1 was described in U.S. Pat. No. 9,499,514 andInternational Patent Publication No. WO 2016/007848, the disclosures ofeach which are incorporated herein by reference in their entireties.

4. SUMMARY

Provided herein are solid forms of2-(4-chlorophenyl)-N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-2,2-difluoroacetamide,and tautomers thereof (Compound 1)

In one embodiment, the solid form is crystalline Form A. In anotherembodiment, the solid form is crystalline Form B. In yet anotherembodiment, the solid form is crystalline Form C. In yet anotherembodiment, the solid form is crystalline Form D. In yet anotherembodiment, the solid form is crystalline Form E.

Further provided herein is an amorphous form comprising Compound 1.

In certain embodiments, the solid forms are single-component crystalforms of2-(4-chlorophenyl)-N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-2,2-difluoroacetamide.In certain embodiments, the solid forms are multiple-component crystalforms of2-(4-chlorophenyl)-N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-2,2-difluoroacetamide.In certain embodiments, the solid forms are single-component amorphousforms of2-(4-chlorophenyl)-N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-2,2-difluoroacetamide.In certain embodiments, the solid forms are multiple-component amorphousforms of2-(4-chlorophenyl)-N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-2,2-difluoroacetamide.Without intending to be limited by any particular theory, certain solidforms provided herein have particular advantageous physical and/orchemical properties making them useful, e.g., for manufacturing,processing, formulation and/or storage, while also possessingparticularly advantageous biological properties, such as, e.g.,bioavailability and/or biological activity.

In certain embodiments, solid forms provided herein include solid formscomprising2-(4-chlorophenyl)-N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-2,2-difluoroacetamide,including, but not limited to, single-component and multiple-componentsolid forms comprising2-(4-chlorophenyl)-N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-2,2-difluoroacetamide.Certain embodiments herein provide methods of making, isolating and/orcharacterizing the solid forms provided herein.

Also provided are pharmaceutical compositions formulated foradministration by an appropriate route and means containing effectiveconcentrations of a solid form of2-(4-chlorophenyl)-N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-2,2-difluoroacetamide,and optionally comprising at least one pharmaceutical carrier.

In one embodiment, the pharmaceutical compositions deliver amountseffective for the treatment of cancer, including solid tumors and bloodborne tumors. In one embodiment, the pharmaceutical compositions deliveramounts effective for the prevention of cancer, including solid tumorsand blood borne tumors. In one embodiment, the pharmaceuticalcompositions deliver amounts effective for the amelioration of cancer,including solid tumors and blood borne tumors.

Also provided herein are combination therapies using a solid form of2-(4-chlorophenyl)-N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-2,2-difluoroacetamidein combination with a therapy e.g., another pharmaceutical agent withactivity against cancer, for example AML, or a myelodysplastic syndrome(MDS), and/or symptoms thereof. Examples of therapies within the scopeof the methods include, but are not limited to, surgery, chemotherapy,radiation therapy, hormonal therapy, biological therapy, immunotherapy,and combinations thereof.

The solid form of2-(4-chlorophenyl)-N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-2,2-difluoroacetamideprovided herein may be administered simultaneously with, prior to, orafter administration of one or more of the above therapies.Pharmaceutical compositions containing a solid form of2-(4-chlorophenyl)-N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-2,2-difluoroacetamideprovided herein and one or more of the above agents are also provided.

In certain embodiments, provided herein are methods of treating,preventing or ameliorating cancer, including solid tumors and bloodborne tumors, or one or more symptoms or causes thereof. In certainembodiments, provided herein are methods of preventing cancer, includingsolid tumors and blood borne tumors, or one or more symptoms or causesthereof. In certain embodiments, provided herein are methods ofameliorating cancer, including solid tumors and blood borne tumors, orone or more symptoms or causes thereof. In certain embodiments, theblood borne tumor is leukemia. In certain embodiments, methods providedherein encompass methods of treating various forms of leukemias such aschronic lymphocytic leukemia, chronic myeloid leukemia, acutelymphocytic leukemia, acute myeloid leukemia and acute myeloblasticleukemia. In certain embodiments, methods provided herein encompassmethods of preventing various forms of leukemias such as chroniclymphocytic leukemia, chronic myeloid leukemia, acute lymphocyticleukemia, acute myeloid leukemia and acute myeloblastic leukemia. Incertain embodiments, methods provided herein encompass methods ofmanaging various forms of leukemias such as chronic lymphocyticleukemia, chronic myeloid leukemia, acute lymphocytic leukemia, acutemyeloid leukemia and acute myeloblastic leukemia. The methods providedherein include treatment of leukemias that are relapsed, refractory orresistant. The methods provided herein include prevention of leukemiasthat are relapsed, refractory or resistant. The methods provided hereininclude management of leukemias that are relapsed, refractory orresistant. In one embodiment, methods provided herein encompass methodsof treating acute myeloid leukemia. In one embodiment, methods providedherein encompass methods of preventing acute myeloid leukemia. In oneembodiment, methods provided herein encompass methods of managing acutemyeloid leukemia.

In certain embodiments, provided herein are methods of treating,preventing or ameliorating a myelodysplastic syndrome, or one or moresymptoms or causes thereof. In certain embodiments, provided herein aremethods of preventing cancer, including a myelodysplastic syndrome, orone or more symptoms or causes thereof. In certain embodiments, providedherein are methods of ameliorating a myelodysplastic syndrome, or one ormore symptoms or causes thereof.

In one embodiment, provided herein are methods of treating acute myeloidleukemia by intravenous administration of a composition comprising solidform of2-(4-chlorophenyl)-N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-2,2-difluoroacetamideor and tautomers thereof.

In one embodiment, provided herein are methods of treating amyelodysplastic syndrome by intravenous administration of a compositioncomprising solid form of2-(4-chlorophenyl)-N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-2,2-difluoroacetamideor and tautomers thereof.

In practicing the methods, effective amounts of a solid form of2-(4-chlorophenyl)-N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-2,2-difluoroacetamideor compositions containing therapeutically effective concentrations ofthe solid form of2-(4-chlorophenyl)-N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-2,2-difluoroacetamideare administered to an individual exhibiting the symptoms of the diseaseor disorder to be treated. The amounts are effective to ameliorate oreliminate one or more symptoms of the disease or disorder.

Further provided is a pharmaceutical pack or kit comprising one or morecontainers filled with one or more of the ingredients of thepharmaceutical compositions. Optionally associated with suchcontainer(s) can be a notice in the form prescribed by a governmentalagency regulating the manufacture, use or sale of pharmaceuticals orbiological products, which notice reflects approval by the agency ofmanufacture, use of sale for human administration. The pack or kit canbe labeled with information regarding mode of administration, sequenceof drug administration (e.g., separately, sequentially or concurrently),or the like.

These and other aspects of the subject matter described herein willbecome evident upon reference to the following detailed description.

5. BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 depicts an X-ray powder diffractogram stack plot of Forms A, B,C, D, and E of Compound 1.

FIG. 2 depicts an X-ray powder diffractogram (XRPD) plot of Form A ofCompound 1.

FIG. 3 depicts a SEM image of Form A of Compound 1.

FIG. 4 depicts a thermogravimetrical analysis (TGA) plot of Form A ofCompound 1.

FIG. 5 depicts a differential scanning calorimetry (DSC) thermogram plotof Form A of Compound 1.

FIG. 6 provides a dynamic vapor sorption (DVS) isotherm plot of Form Aof Compound 1.

FIG. 7 provides a ¹H NMR spectrum of Form A of Compound 1.

FIG. 8 depicts the comparison of the X-ray powder diffractogram plots ofForm A of Compound 1 before (a) and after (b) compression.

FIG. 9 depicts an XRPD plot of Form B of Compound 1.

FIG. 10 depicts a SEM image of Form B of Compound 1.

FIG. 11 depicts a TGA thermogram plot of Form B of Compound 1.

FIG. 12 depicts a DSC thermogram plot of Form B of Compound 1.

FIG. 13 provides a DVS isothterm plot of Form B of Compound 1.

FIG. 14 provides a ¹H NMR spectrum of Form B of Compound 1.

FIG. 15 depicts the comparison of the X-ray powder diffractogram plotsof Form B of Compound 1 before (a) and after (b) compression.

FIG. 16 depicts an XRPD plot of Form C of Compound 1.

FIG. 17 depicts a SEM image of Form C of Compound 1.

FIG. 18 depicts a TGA thermogram plot of Form C of Compound 1.

FIG. 19 depicts a DSC thermogram of Form C of Compound 1.

FIG. 20 provides a DVS isotherm plot of Form C of Compound 1.

FIG. 21 provides a ¹H NMR spectrum of Form C of Compound 1.

FIG. 22 depicts the comparison of the X-ray powder diffractogram plotsof Form C of Compound 1 before (a) and after (b) compression.

FIG. 23 depicts an XRPD plot of Form D of Compound 1.

FIG. 24 depicts a TGA thermogram plot of Form D of Compound 1.

FIG. 25 depicts an XRPD plot of Form E of Compound 1.

FIG. 26 depicts a TGA thermogram plot of Form E of Compound 1.

FIG. 27 depicts the modulated DSC thermogramplot of amorphous Compound1.

FIG. 28 depicts an XRPD plot of amorphous Compound 1.

FIG. 29 depicts a ¹H NMR spectrum of amorphous Compound 1.

6. DETAILED DESCRIPTION 6.1. Definitions

As used herein, the term “Compound 1” refers to2-(4-chlorophenyl)-N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-2,2-difluoroacetamide,which has the following structure:

and tautomers thereof.

As used herein and unless otherwise specified, the terms “solid form”and related terms refer to a physical form which is not predominantly ina liquid or a gaseous state. As used herein and unless otherwisespecified, the term “solid form” and related terms, when used herein torefer to Compound 1, refer to a physical form comprising Compound 1which is not predominantly in a liquid or a gaseous state. The solidforms may be crystalline, amorphous, or mixtures thereof. In particularembodiments, solid forms may be liquid crystals. A “single-component”solid form comprising Compound 1 consists essentially of Compound 1. A“multiple-component” solid form comprising Compound 1 comprises asignificant quantity of one or more additional species, such as ionsand/or molecules, within the solid form. Multiple-component solid formscomprising Compound 1 include co-crystals, solvates (e.g., hydrates),and clathrates of Compound 1. In particular embodiments, the term “solidform comprising Compound 1” and related terms include single-componentand multiple-component solid forms comprising Compound 1. In particularembodiments, “solid forms comprising Compound 1” and related termsinclude crystal forms comprising Compound 1, amorphous forms comprisingCompound 1, and mixtures thereof.

As used herein and unless otherwise specified, the term “crystalline”and related terms used herein, when used to describe a compound,substance, modification, material, component or product, unlessotherwise specified, mean that the compound, substance, modification,material, component or product is substantially crystalline asdetermined by X-ray diffraction. See, e.g., Remington: The Science andPractice of Pharmacy, 21^(st) edition, Lippincott, Williams and Wilkins,Baltimore, Md. (2005); The United States Pharmacopeia, 23^(rd) ed.,1843-1844 (1995).

As used herein and unless otherwise specified, the term “crystal forms,”“crystalline forms” and related terms herein refer to solid forms thatare crystalline. Crystal forms include single-component crystal formsand multiple-component crystal forms, and include polymorphs, solvates,hydrates, and/or other molecular complexes. In certain embodiments, acrystal form of a substance may be substantially free of amorphous formsand/or other crystal forms. In certain embodiments, a crystal form of asubstance may contain less than about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%,9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45% or 50% of one or moreamorphous forms and/or other crystal forms on a weight basis. In certainembodiments, a crystal form of a substance may be physically and/orchemically pure. In certain embodiments, a crystal form of a substancemay be about 99%, 98%, 97%, 96%, 95%, 94%, 93%, 92%, 91% or 90%physically and/or chemically pure.

As used herein and unless otherwise specified, the terms “polymorphs,”“polymorphic forms” and related terms herein, refer to two or morecrystal forms that consist essentially of the same molecule, molecules,and/or ions. Like different crystal forms, different polymorphs may havedifferent physical properties such as, e.g., melting temperature, heatof fusion, solubility, dissolution properties and/or vibrationalspectra, as a result of the arrangement or conformation of the moleculesand/or ions in the crystal lattice. The differences in physicalproperties may affect pharmaceutical parameters such as storagestability, compressibility and density (important in formulation andproduct manufacturing), and dissolution rate (an important factor inbioavailability). Differences in stability can result from changes inchemical reactivity (e.g., differential oxidation, such that a dosageform discolors more rapidly when comprised of one polymorph than whencomprised of another polymorph) or mechanical changes (e.g., tabletscrumble on storage as a kinetically favored polymorph converts tothermodynamically more stable polymorph) or both (e.g., tablets of onepolymorph are more susceptible to breakdown at high humidity). As aresult of solubility/dissolution differences, in the extreme case, somesolid-state transitions may result in lack of potency or, at the otherextreme, toxicity. In addition, the physical properties may be importantin processing (e.g., one polymorph might be more likely to form solvatesor might be difficult to filter and wash free of impurities, andparticle shape and size distribution might be different betweenpolymorphs).

As used herein and unless otherwise specified, the terms “solvate” and“solvated,” refer to a crystal form of a substance which containssolvent. The terms “hydrate” and “hydrated” refer to a solvate whereinthe solvent comprises water. “Polymorphs of solvates” refers to theexistence of more than one crystal form for a particular solvatecomposition. Similarly, “polymorphs of hydrates” refers to the existenceof more than one crystal form for a particular hydrate composition. Theterm “desolvated solvate,” as used herein, refers to a crystal form of asubstance which may be prepared by removing the solvent from a solvate.

As used herein and unless otherwise specified, the term “amorphous,”“amorphous form,” and related terms used herein, mean that thesubstance, component or product in question is not substantiallycrystalline as determined by X-ray diffraction. In particular, the term“amorphous form” describes a disordered solid form, i.e., a solid formlacking long range crystalline order. In certain embodiments, anamorphous form of a substance may be substantially free of otheramorphous forms and/or crystal forms. In other embodiments, an amorphousform of a substance may contain less than about 1%, 2%, 3%, 4%, 5%, 10%,15%, 20%, 25%, 30%, 35%, 40%, 45% or 50% of one or more other amorphousforms and/or crystal forms on a weight basis. In certain embodiments, anamorphous form of a substance may be physically and/or chemically pure.In certain embodiments, an amorphous form of a substance be about 99%,98%, 97%, 96%, 95%, 94%, 93%, 92%, 91% or 90% physically and/orchemically pure.

“Tautomers” refers to isomeric forms of a compound that are inequilibrium with each other. The concentrations of the isomeric formswill depend on the environment the compound is found in and may bedifferent depending upon, for example, whether the compound is a solidor is in an organic or aqueous solution. For example, in aqueoussolution, pyrazoles may exhibit the following isomeric forms, which arereferred to as tautomers of each other:

As readily understood by one skilled in the art, a wide variety offunctional groups and other structures may exhibit tautomerism and alltautomers of Compound 1 are within the scope of the present invention.

Techniques for characterizing crystal forms and amorphous forms include,but are not limited to, thermal gravimetric analysis (TGA), differentialscanning calorimetry (DSC), X-ray powder diffractometry (XRPD),single-crystal X-ray diffractometry, vibrational spectroscopy, e.g.,infrared (IR) and Raman spectroscopy, solid-state and solution nuclearmagnetic resonance (NMR) spectroscopy, optical microscopy, hot stageoptical microscopy, scanning electron microscopy (SEM), electroncrystallography and quantitative analysis, particle size analysis (PSA),surface area analysis, solubility measurements, dissolutionmeasurements, elemental analysis, and Karl Fischer analysis.Characteristic unit cell parameters may be determined using one or moretechniques such as, but not limited to, X-ray diffraction and neutrondiffraction, including single-crystal diffraction and powderdiffraction. Techniques useful for analyzing powder diffraction datainclude profile refinement, such as Rietveld refinement, which may beused, e.g., to analyze diffraction peaks associated with a single phasein a sample comprising more than one solid phase. Other methods usefulfor analyzing powder diffraction data include unit cell indexing, whichallows one of skill in the art to determine unit cell parameters from asample comprising crystalline powder.

As used herein and unless otherwise specified, the terms “about” and“approximately,” when used in connection with a numeric value or a rangeof values which is provided to characterize a particular solid form,e.g., a specific temperature or temperature range, such as, e.g., thatdescribing a DSC or TGA thermal event, including, e.g., melting,dehydration, desolvation or glass transition events; a mass change, suchas, e.g., a mass change as a function of temperature or humidity; asolvent or water content, in terms of, e.g., mass or a percentage; or apeak position, such as, e.g., in analysis by IR or Raman spectroscopyor)(RFD; indicate that the value or range of values may deviate to anextent deemed reasonable to one of ordinary skill in the art while stilldescribing the particular solid form. For example, in particularembodiments, the terms “about” and “approximately,” when used in thiscontext and unless otherwise specified, indicate that the numeric valueor range of values may vary within 25%, 20%, 15%, 10%, 9%, 8%, 7%, 6%,5%, 4%, 3%, 2%, 1.5%, 1%, 0.5%, or 0.25% of the recited value or rangeof values.

As used herein and unless otherwise specified, a sample comprising aparticular crystal form or amorphous form that is “substantially pure,”e.g., substantially free of other solid forms and/or of other chemicalcompounds, contains, in particular embodiments, less than about 25%,20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.75%, 0.5%, 0.25% or0.1% percent by weight of one or more other solid forms and/or of otherchemical compounds.

As used herein and unless otherwise specified, a sample or compositionthat is “substantially free” of one or more other solid forms and/orother chemical compounds means that the composition contains, inparticular embodiments, less than about 25%, 20%, 15%, 10%, 9%, 8%, 7%,6%, 5%, 4%, 3%, 2%, 1%, 0.75%, 0.5%, 0.25% or 0.1% percent by weight ofone or more other solid forms and/or other chemical compounds.

As used herein, the term “patient” refers to a mammal, particularly ahuman.

As used herein, term “adverse effects” includes, but is not limited togastrointestinal, renal and hepatic toxicities, leukopenia, increases inbleeding times due to, e.g., thrombocytopenia, and prolongation ofgestation, nausea, vomiting, somnolence, asthenia, dizziness,teratogenicity, extra-pyramidal symptoms, akathisia, cardiotoxicityincluding cardiovascular disturbances, inflammation, male sexualdysfunction, and elevated serum liver enzyme levels. The term“gastrointestinal toxicities” includes, but is not limited to, gastricand intestinal ulcerations and erosions. The term “renal toxicities”includes, but is not limited to, such conditions as papillary necrosisand chronic interstitial nephritis.

As used herein and unless otherwise indicated, the phrases “reduce oravoid adverse effects” and “reducing or avoiding adverse effects” meanthe reduction of the severity of one or more adverse effects as definedherein.

As used herein, and unless otherwise specified, the terms “treat,”“treating” and “treatment” refer to the eradication or amelioration of adisease or disorder, or of one or more symptoms associated with thedisease or disorder. In certain embodiments, the terms refer tominimizing the spread or worsening of the disease or disorder resultingfrom the administration of one or more prophylactic or therapeuticagents to a patient with such a disease or disorder. In someembodiments, the terms refer to the administration of a compoundprovided herein, with or without other additional active agent, afterthe onset of symptoms of the particular disease.

As used herein, and unless otherwise specified, the terms “prevent,”“preventing” and “prevention” refer to the prevention of the onset,recurrence or spread of a disease or disorder, or of one or moresymptoms thereof. In certain embodiments, the terms refer to thetreatment with or administration of a compound provided herein, with orwithout other additional active compound, prior to the onset ofsymptoms, particularly to patients at risk of diseases or disordersprovided herein. The terms encompass the inhibition or reduction of asymptom of the particular disease. Patients with familial history of adisease in particular are candidates for preventive regimens in certainembodiments. In addition, patients who have a history of recurringsymptoms are also potential candidates for the prevention. In thisregard, the term “prevention” may be interchangeably used with the term“prophylactic treatment.”

As used herein, and unless otherwise specified, the terms “manage,”“managing” and “management” refer to preventing or slowing theprogression, spread or worsening of a disease or disorder, or of one ormore symptoms thereof. Often, the beneficial effects that a patientderives from a prophylactic and/or therapeutic agent do not result in acure of the disease or disorder. In this regard, the term “managing”encompasses treating a patient who had suffered from the particulardisease in an attempt to prevent or minimize the recurrence of thedisease.

As used herein, and unless otherwise specified, a “therapeuticallyeffective amount” of a compound is an amount sufficient to provide atherapeutic benefit in the treatment and/or management of a disease ordisorder, or to delay or minimize one or more symptoms associated withthe disease or disorder. A therapeutically effective amount of acompound means an amount of therapeutic agent, alone or in combinationwith other therapies, which provides a therapeutic benefit in thetreatment or management of the disease or disorder. The term“therapeutically effective amount” can encompass an amount that improvesoverall therapy, reduces or avoids symptoms or causes of disease ordisorder, or enhances the therapeutic efficacy of another therapeuticagent.

As used herein, and unless otherwise specified, a “prophylacticallyeffective amount” of a compound is an amount sufficient to prevent adisease or disorder, or prevent its recurrence. A prophylacticallyeffective amount of a compound means an amount of therapeutic agent,alone or in combination with other agents, which provides a prophylacticbenefit in the prevention of the disease. The term “prophylacticallyeffective amount” can encompass an amount that improves overallprophylaxis or enhances the prophylactic efficacy of anotherprophylactic agent.

As used herein, the term “tumor,” refers to all neoplastic cell growthand proliferation, whether malignant or benign, and all pre-cancerousand cancerous cells and tissues. “Neoplastic,” as used herein, refers toany form of dysregulated or unregulated cell growth, whether malignantor benign, resulting in abnormal tissue growth. Thus, “neoplastic cells”include malignant and benign cells having dysregulated or unregulatedcell growth.

As used herein, “hematologic malignancy” refers to cancer of the body'sblood-forming and immune system-the bone marrow and lymphatic tissue.Such cancers include leukemias, lymphomas (Non-Hodgkin's Lymphoma),Hodgkin's disease (also called Hodgkin's Lymphoma) and myeloma. In oneembodiment, the myeloma is multiple myeloma. In some embodiments, theleukemia is, for example, acute myelogenous leukemia (AML), acutelymphocytic leukemia (ALL), adult T-cell leukemia, chronic lymphocyticleukemia (CLL), hairy cell leukemia, myelodysplasia, myeloproliferativedisorders, chronic myelogenous leukemia (CML), myelodysplastic syndrome(MDS), human lymphotropic virus-type 1 (HTLV-1) leukemia, mastocytosis,or B-cell acute lymphoblastic leukemia. In some embodiments, thelymphoma is, for example, diffuse large B-cell lymphoma (DLBCL), B-cellimmunoblastic lymphoma, small non-cleaved cell lymphoma, humanlymphotropic virus-type 1 (HTLV-1) leukemia/lymphoma, adult T-celllymphoma, peripheral T-cell lymphoma (PTCL), cutaneous T-cell lymphoma(CTCL), mantle cell lymphoma (MCL), Hodgkin lymphoma (HL), non-Hodgkinlymphoma (NHL), AIDS-related lymphoma, follicular lymphoma, smalllymphocytic lymphoma, T-cell/histiocyte rich large B-cell lymphoma,transformed lymphoma, primary mediastinal (thymic) large B-celllymphoma, splenic marginal zone lymphoma, Richter's transformation,nodal marginal zone lymphoma, or ALK-positive large B-cell lymphoma. Inone embodiment, the hematological cancer is indolent lymphoma including,for example, DLBCL, follicular lymphoma, or marginal zone lymphoma.

The term “leukemia” refers to malignant neoplasms of the blood-formingtissues. The leukemia includes, but is not limited to, chroniclymphocytic leukemia, chronic myelocytic leukemia, acute lymphoblasticleukemia, acute myeloid leukemia, and acute myeloblastic leukemia. Theleukemia can be relapsed, refractory or resistant to conventionaltherapy.

The term “myelodysplastic syndrome” refers to hematological conditionscharacterized by abnormalities in the production of one or more of thecellular components of blood (red cells, white cells (other thanlymphocytes) and platelets (or their progenitor cells, megakaryocytes)),and includes the following disorders: refractory anemia (RA); RA withringed sideroblasts (RARS); RA with excess of blasts (RAEB); refractorycytopenia with multilineage dysplasia (RCMD), refractory cytopenia withunilineage dysplasia (RCUD); unclassifiable myelodysplastic syndrome(MDS-U), myelodysplastic syndrome associated with an isolated del(5q)chromosome abnormality, therapy-related myeloid neoplasms and chronicmyelomonocytic leukemia (CMML).

As used herein, “promyelocytic leukemia” or “acute promyelocyticleukemia” refers to a malignancy of the bone marrow in which there is adeficiency of mature blood cells in the myeloid line of cells and anexcess of immature cells called promyelocytes. It is usually marked byan exchange of regions of chromosomes 15 and 17.

As used herein, “acute lymphocytic leukemia (ALL)”, also known as “acutelymphoblastic leukemia” refers to a malignant disease caused by theabnormal growth and development of early nongranular white blood cells,or lymphocytes.

As used herein, “T-cell leukemia” refers to a disease in which certaincells of the lymphoid system called T lymphocytes or T cells aremalignant. T cells are white blood cells that normally can attackvirus-infected cells, foreign cells, and cancer cells and producesubstances that regulate the immune response.

The term “relapsed” refers to a situation where patients who have had aremission of leukemia after therapy have a return of leukemia cells inthe marrow and a decrease in normal blood cells.

The term “refractory or resistant” refers to a circumstance wherepatients, even after intensive treatment, have residual leukemia cellsin their marrow.

The terms “co-administration” and “in combination with” include theadministration of two therapeutic agents (for example, a compoundprovided herein and another anti-cancer agent) either simultaneously,concurrently or sequentially with no specific time limits. In oneembodiment, both agents are present in the cell or in the patient's bodyat the same time or exert their biological or therapeutic effect at thesame time. In one embodiment, the two therapeutic agents are in the samecomposition or unit dosage form. In another embodiment, the twotherapeutic agents are in separate compositions or unit dosage forms.

The term “the supportive care agent” refers to any substance thattreats, prevents and/or manages an adverse effect from treatment withthe solid form of Compound 1.

The term “biological therapy” refers to administration of biologicaltherapeutics such as cord blood, stem cells, growth factors and thelike.

As used herein, overall survival (OS) means the time from randomizationin a clinical trial until death from any cause. As used herein,progression-free survival (PFS) means the time from randomization in aclinical trial until progression or death. As used herein, event-freesurvival (EFS) means the time from study entry until any treatmentfailure, including disease progression, treatment discontinuation forany reason, or death. As used herein, overall response rate (ORR) meansthe sum of the percentage of patients who achieve complete and partialresponsess. As used herein, duration of response (DoR) is the time fromachieving a response until relapse or disease progression.

“Anti-cancer agents” refer to anti-metabolites (e.g., 5-fluoro-uracil,methotrexate, fludarabine), antimicrotubule agents (e.g., vincaalkaloids such as vincristine, vinblastine; taxanes such as paclitaxel,docetaxel), alkylating agents (e.g., cyclophosphamide, melphalan,carmustine, nitrosoureas such as bischloroethylnitrosurea andhydroxyurea), platinum agents (e.g. cisplatin, carboplatin, oxaliplatin,JM-216 or satraplatin, CI-973), anthracyclines (e.g., doxorubicin,daunorubicin), antitumor antibiotics (e.g., mitomycin, idarubicin,adriamycin, daunomycin), topoisomerase inhibitors (e.g., etoposide,camptothecins), anti-angiogenesis agents (e.g. Sutent® and Bevacizumab)or any other cytotoxic agents, (estramustine phosphate, prednimustine),hormones or hormone agonists, antagonists, partial agonists or partialantagonists, kinase inhibitors, checkpoint inhibitors, and radiationtreatment.

The term “composition” as used herein is intended to encompass a productcomprising the specified ingredients (and in the specified amounts, ifindicated), as well as any product which results, directly orindirectly, from combination of the specified ingredients in thespecified amounts. By “pharmaceutically acceptable” it is meant that thediluent, excipient or carrier must be compatible with the otheringredients of the formulation and not deleterious to the recipientthereof.

It should be noted that if there is a discrepancy between a depictedstructure and a name given that structure, the depicted structure is tobe accorded more weight. In addition, if the stereochemistry of astructure or a portion of a structure is not indicated with, forexample, bold or dashed lines, the structure or portion of the structureis to be interpreted as encompassing all stereoisomers of it.

This disclosure relates to solid forms of Compound 1, which is2-(4-chlorophenyl)-N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-2,2-difluoroacetamideand tautomers thereof, as well as methods of using, and compositionscomprising, a solid form of Compound 1. For example, the presentdisclosure encompasses the in vitro and in vivo use of a solid form ofCompound 1, and the incorporation of a solid form of Compound 1 intopharmaceutical compositions and single unit dosage forms useful in thetreatment and prevention of a variety of diseases and disorders.

6.2. Solid Forms of Compound 1

In one embodiment, provided herein are solid forms of Compound 1.

Compound 1 can be prepared using the methods as described in U.S. Pat.No. 9,499,514 and International Patent Publication No. WO 2016/007848,the disclosures of each which are incorporated herein by reference intheir entireties.

The solid forms comprising Compound 1 include single-component andmultiple-component forms, including crystal forms and amorphous forms.Particular embodiments herein provide single-component amorphous solidforms of Compound 1. Particular embodiments herein providesingle-component crystalline solid forms of Compound 1. Particularembodiments herein provide multiple-component amorphous forms comprisingCompound 1. Particular embodiments herein provide multiple-componentcrystalline solid forms comprising Compound 1.

The solid forms comprising Compound 1 can be prepared by the methodsdescribed herein, including the methods described in the Examples below,or by techniques known in the art, including heating, cooling, freezedrying, lyophilization, quench cooling the melt, rapid solventevaporation, slow solvent evaporation, solvent recrystallization,antisolvent addition, slurry recrystallization, crystallization from themelt, desolvation, recrystallization in confined spaces such as, e.g.,in nanopores or capillaries, recrystallization on surfaces or templatessuch as, e.g., on polymers, recrystallization in the presence ofadditives, such as, e.g., co-crystal counter-molecules, desolvation,dehydration, rapid cooling, slow cooling, exposure to solvent and/orwater, drying, including, e.g., vacuum drying, vapor diffusion,sublimation, grinding (including, e.g., cryo-grinding, solvent-dropgrinding or liquid assisted grinding), microwave-induced precipitation,sonication-induced precipitation, laser-induced precipitation andprecipitation from a supercritical fluid. The particle size of theresulting solid forms, which can vary, (e.g., from nanometer dimensionsto millimeter dimensions), can be controlled, e.g., by varyingcrystallization conditions, such as, e.g., the rate of crystallizationand/or the crystallization solvent system, or by particle-size reductiontechniques, e.g., grinding, milling, micronizing or sonication.

While not intending to be bound by any particular theory, certain solidforms are characterized by physical properties, e.g., stability,solubility and dissolution rate, appropriate for pharmaceutical andtherapeutic dosage forms. Moreover, while not wishing to be bound by anyparticular theory, certain solid forms are characterized by physicalproperties (e.g., density, compressibility, hardness, morphology,cleavage, stickiness, solubility, water uptake, electrical properties,thermal behavior, solid-state reactivity, physical stability, andchemical stability) affecting particular processes (e.g., yield,filtration, washing, drying, milling, mixing, tableting, flowability,dissolution, formulation, and lyophilization) which make certain solidforms suitable for the manufacture of a solid dosage form. Suchproperties can be determined using particular analytical chemicaltechniques, including solid-state analytical techniques (e.g., X-raydiffraction, microscopy, spectroscopy and thermal analysis), asdescribed herein and known in the art.

The solid forms provided herein (e.g., Form A, Form B, Form C, Form D,Form E and amorphous of Compound 1) may be characterized using a numberof methods known to a person having ordinary skill in the art,including, but not limited to, single crystal X-ray diffraction, X-raypowder diffraction (MUD), microscopy (e.g., scanning electron microscopy(SEM)), thermal analysis (e.g., differential scanning calorimetry (DSC),thermal gravimetric analysis (TGA), and hot-stage microscopy),spectroscopy (e.g., infrared, Raman, and solid-state nuclear magneticresonance), single differential thermal analysis (SDTA), highperformance liquid chromatography coupled with mass spectroscopy(HPLC-MS), thermogravimetrical analysis coupled with single differentialthermal analysis (TGA-SDTA), and thermogravimetric analysis coupled withmass spectroscopy (TGA-MS). The particle size and size distribution ofthe solid form provided herein may be determined by conventionalmethods, such as laser light scattering technique.

The purity of the solid forms provided herein may be determined bystandard analytical methods, such as thin layer chromatography (TLC),gel electrophoresis, gas chromatography, high performance liquidchromatography (HPLC), and mass spectrometry (MS).

It should be understood that the numerical values of the peaks of anX-ray powder diffraction pattern may vary slightly from one machine toanother or from one sample to another, and so the values quoted are notto be construed as absolute, but with an allowable variability, such as±0.2 degrees two theta (° 2θ) (see United State Pharmacopoeia, page 2228(2003)).

Certain embodiments herein provide compositions comprising one or moreof the solid forms. Certain embodiments provide compositions of one ormore solid forms in combination with other active ingredients. Certainembodiments provide methods of using these compositions in thetreatment, prevention and/or management of diseases and disordersincluding, but not limited to, the diseases and disorders providedherein.

The solid forms provided herein may also comprise unnatural proportionsof atomic isotopes at one or more of the atoms in Compound 1. Forexample, the compound may be radiolabeled with radioactive isotopes,such as for example tritium (³H), or carbon-14 (¹⁴C), or may beisotopically enriched, such as with deuterium (²H), carbon-13 (¹³C), ornitrogen-15 (¹⁵N). As used herein, an “isotopologue” is an isotopicallyenriched compound. The term “isotopically enriched” refers to an atomhaving an isotopic composition other than the natural isotopiccomposition of that atom. “Isotopically enriched” may also refer to acompound containing at least one atom having an isotopic compositionother than the natural isotopic composition of that atom. The term“isotopic composition” refers to the amount of each isotope present fora given atom. Radiolabeled and isotopically encriched compounds areuseful as therapeutic agents, e.g., cancer and inflammation therapeuticagents, research reagents, e.g., binding assay reagents, and diagnosticagents, e.g., in vivo imaging agents. All isotopic variations ofCompound 1, whether radioactive or not, are intended to be encompassedwithin the scope of the embodiments provided herein. In someembodiments, there are provided isotopologues of Compound 1, forexample, the isotopologues are deuterium, carbon-13, or nitrogen-15enriched Compound 1.

The compound may be radiolabeled with radioactive isotopes, such as forexample tritium (³H), iodine-125 (¹²⁵I) sulfur-35 (³⁵S), or carbon-14(¹⁴C). Radiolabeled compounds are useful as therapeutic agents, e.g.,cancer therapeutic agents, research reagents, e.g., binding assayreagents, and diagnostic agents, e.g., in vivo imaging agents. Allisotopic variations of Compound 1 or isotopologies of Compound 1,whether radioactive or not, are intended to be encompassed within thescope of the embodiments provided herein.

6.2.1. Form a of Compound 1

In certain embodiments, provided herein is Form A of Compound 1.

In one embodiment, Form A is an anhydrous form of Compound 1. In anotherembodiment, Form A of Compound 1 is crystalline.

In certain embodiments, Form A is obtained by crystallization fromcertain solvent systems, for example, solvent systems comprising one ormore of the following solvents: acetone and the solvent mixture ofisopropanol and water at room temperature. In certain embodiments, FormA is obtained as an intermediate solid form from slurries at elevatedtemperature, for example about 50° C., in ethanol/water (1:1), acetoneor acetonitrile.

In certain embodiments, Form A is substantially crystalline, asindicated by, e.g., X-ray powder diffraction measurements. In oneembodiment, Form A of Compound 1 has an X-ray powder diffraction patternsubstantially as shown in FIG. 2.

In one embodiment, Form A of Compound 1 has one or more characteristicX-ray powder diffraction peaks at a two-theta angle of approximately11.5, 15.6, 16.6, 17.2, 18.1, 19.0, 19.6, 21.1, 23.2 or 24.8 degrees 2θas depicted in FIG. 2. In another embodiment, Form A of Compound 1 hasone, two, three or four characteristic X-ray powder diffraction peaks ata two-theta angle of approximately 15.6, 16.6, 17.2 or 24.8 degrees 2θ.In another embodiment, Form A of Compound 1 has one, two, three, four,five, six or seven characteristic X-ray powder diffraction peaks as setforth in Table 1. In another embodiment, Form A of Compound 1 has one,two, or three characteristic X-ray powder diffraction peaks as set forthin Table 1.

In one embodiment, Form A of Compound 1 has the SEM picture as shown inFIG. 3.

In one embodiment, provided herein is a crystalline form of Compound 1having a thermogravimetric (TGA) thermograph corresponding substantiallyto the representative TGA thermogram as depicted in FIG. 4. In certainembodiments, no TGA weight loss is observed for Form A.

In one embodiment, provided herein is crystalline form A of Compound 1having a DSC thermogram corresponding substantially as depicted in FIG.5. In certain embodiments, Form A is characterized by a DSC plotcomprising a melting event with an onset temperature of 229° C. and heatof fusion of 118 J/g.

In certain embodiments, Form A is characterized by dynamic vaporsorption analysis. A representative dynamic vapor sorption (DVS)isotherm plot is shown in FIG. 6. In certain embodiments, when therelative humidity (“RH”) is increased from about 0% to about 90% RH,Form A exhibits less than 1.5%, less than 1.2% or about 1.2% w/w wateruptake. In certain embodiments, Form A comprises less than 0.1% water asdetermined in a coulometric Karl Fischer (KF) titrator equipped with anoven sample processor set at 225° C.

In certain embodiments, no significant degradation or residual solventfor Form A is observed by ¹H NMR (FIG. 7).

In certain embodiments, Form A of Compound 1 is characterized by itsstability profile upon compression. In certain embodiments, Form A isstable, e.g., its XRPD pattern remains substantially unchanged withbroader diffraction peaks, upon application of 2000-psi pressure forabout 1 minute (FIG. 8).

In still another embodiment, Form A of Compound 1 is substantially pure.In certain embodiments, the substantially pure Form A of Compound 1 issubstantially free of other solid forms, e.g., amorphous form. Incertain embodiments, the purity of the substantially pure Form A ofCompound 1 is no less than about 95% pure, no less than about 96% pure,no less than about 97% pure, no less than about 98% pure, no less thanabout 98.5% pure, no less than about 99% pure, no less than about 99.5%pure, or no less than about 99.8% pure.

Certain embodiments herein provide Form A of Compound 1 which issubstantially pure. Certain embodiments herein provide Form A ofCompound 1 which is substantially free of other solid forms comprisingCompound 1 including, e.g., Forms B, C, D, E and/or an amorphous solidform comprising Compound 1 as provided herein. Certain embodimentsherein provide Form A as a mixture of solid forms comprising Compound 1,including, e.g., a mixture comprising one or more of the following:Forms B, C, D, E and an amorphous solid form comprising Compound 1 asprovided herein.

6.2.2. Form B of Compound 1

In certain embodiments, provided herein is anhydrous Form B of Compound1.

In certain embodiments, Form B is obtained by anti-solventrecrystallization from certain solvent systems, for example, solventsystems comprising one or more of the following solvents:methanol/water, DMSO/isopropanol, DMSO/toluene, and DMSO/water. Incertain embodiments, Form B is obtained by cooling recrystallizationfrom THF/water (1:1).

In certain embodiments, Form B is crystalline, as indicated by, e.g.,X-ray powder diffraction measurements. In one embodiment, Form B ofCompound 1 has an X-ray powder diffraction pattern substantially asshown in FIG. 9.

In one embodiment, Form B of Compound 1 has one or more characteristicX-ray powder diffraction peaks at a two-theta angle of approximately15.4, 16.3, 16.7, 17.7, 20.4, 25.6 or 27.5, degrees 2θ as depicted inFIG. 9. In another embodiment, Form B of Compound 1 has one, two, threeor four characteristic X-ray powder diffraction peaks at a two-thetaangle of approximately 16.7, 25.6, 15.4 or 16.3 degrees 2θ. In anotherembodiment, Form B of Compound 1 has one, two, three, four, five, six orseven characteristic X-ray powder diffraction peaks as set forth inTable 2. In another embodiment, Form B of Compound 1 has one, two, orthree characteristic X-ray powder diffraction peaks as set forth inTable 2.

In one embodiment, Form B of Compound 1 has the SEM picture as shown inFIG. 10. In one embodiment, provided herein is a crystalline form ofCompound 1 having a thermogravimetric (TGA) thermograph correspondingsubstantially to the representative TGA thermogram as depicted in FIG.11. In certain embodiments, Form B shows no TGA weight loss below 170°C. In certain embodiments, Form B shows a TGA weight loss of 0.4%between 170˜230° C.

In one embodiment, provided herein is crystalline Form B of Compound 1having a DSC thermogram corresponding substantially as depicted in FIG.12. In certain embodiments, Form B is characterized by a DSC plotcomprising a melt/recrystallization event at 219˜224° C. and a majormelting event with a peak temperature of 231° C.

In certain embodiments, Form B is characterized by dynamic vaporsorption analysis. A representative dynamic vapor sorption (DVS)isotherm plot is shown in FIG. 13. In certain embodiments, when therelative humidity (“RH”) is increased from about 0% to about 90% RH,Form B exhibits about 1.4% w/w water uptake. In certain embodiments,Form B comprises less than 0.1% water as determined in a coulometricKarl Fischer (KF) titrator equipped with an oven sample processor set at225° C.

In certain embodiments, Form B shows no significant degradation orresidual solvent by ¹H NMR (FIG. 14).

In certain embodiments, Form B of Compound 1 is characterized by itsstability profile upon compression. In certain embodiments, Form B isstable, e.g., its XRPD pattern remains substantially unchanged withbroader diffraction peaks, upon application of 2000-psi pressure forabout 1 minute (FIG. 15).

In still another embodiment, Form B of Compound 1 is substantially pure.In certain embodiments, the substantially pure Form B of Compound 1 issubstantially free of other solid forms, e.g., amorphous form. Incertain embodiments, the purity of the substantially pure Form B ofCompound 1 is no less than about 95% pure, no less than about 96% pure,no less than about 97% pure, no less than about 98% pure, no less thanabout 98.5% pure, no less than about 99% pure, no less than about 99.5%pure, or no less than about 99.8% pure.

Certain embodiments herein provide Form B of Compound 1 which issubstantially pure. Certain embodiments herein provide Form B ofCompound 1 which is substantially free of other solid forms comprisingCompound 1 including, e.g., Forms A, C, D, E, and/or an amorphous solidform comprising Compound 1 as provided herein. Certain embodimentsherein provide Form B as a mixture of solid forms comprising Compound 1,including, e.g., a mixture comprising one or more of the following:Forms A, C, D, E, and an amorphous solid form comprising Compound 1 asprovided herein.

6.2.3. Form C of Compound 1

In certain embodiments, provided herein is anhydrous Form C ofCompound 1. In certain embodiments, Form C is the most thermodynamicallystable anhydrate among the crystal forms of Compound 1 provided herein.

In certain embodiments, Form C is obtained by slurrying Compound 1 incertain solvent systems, for example, solvent systems comprising one ormore of the following solvents: acetonitrile/water, acetone, orethanol/water for an extended period of time.

In certain aspects, Form C is obtained by slurrying Form B (1× wt) inacetone (30×vol) at an elevated temperature, for example, from 60-80° C.or 70-75° C. for at least 24 hours, and cooling the mixture to roomtemperature. In one aspect, the slurrying is conducted at a temperatureof 70-75° C. under nitrogen pressure of 50-55-psi. In one aspect, themixture is cooled to room temperature over at least 6 hours.

In certain embodiments, Form C is crystalline, as indicated by, e.g.,X-ray powder diffraction measurements. In one embodiment, Form C ofCompound 1 has an X-ray powder diffraction pattern substantially asshown in FIG. 16.

In one embodiment, Form C of Compound 1 has one or more characteristicX-ray powder diffraction peaks at a two-theta angle of approximately7.4, 11.5, 15.8, 16.7, 16.9, 17.7, 18.4, 19.2, 19.5, 21.1, 23.4, 24.7,or 29.9, degrees 2θ as depicted in FIG. 16. In another embodiment, FormC of Compound 1 has one, two, three or four characteristic X-ray powderdiffraction peaks at a two-theta angle of approximately 16.7, 16.9, 17.7or 24.7 degrees 2θ. In another embodiment, Form C of Compound 1 has one,two, three, four, five, six or seven characteristic X-ray powderdiffraction peaks as set forth in Table 3. In another embodiment, Form Cof Compound 1 has one, two, or three characteristic X-ray powderdiffraction peaks as set forth in Table 3.

In one embodiment, Form C of Compound 1 has the SEM picture as shown inFIG. 17. In one embodiment, provided herein is a crystalline form ofCompound 1 having a thermogravimetric (TGA) thermograph correspondingsubstantially to the representative TGA thermogram as depicted in FIG.18. In certain embodiments, Form C shows no TGA weight loss.

In one embodiment, provided herein is crystalline Form C of Compound 1having a DSC thermogram corresponding substantially as depicted in FIG.19. In certain embodiments, Form C is characterized by a DSC plotcomprising melting event with an onset temperature of 232° C. and heatof fusion of 126 J/g.

In certain embodiments, Form C is characterized by dynamic vaporsorption analysis. A representative dynamic vapor sorption (DVS)isotherm plot is shown in FIG. 20. In certain embodiments, when therelative humidity (“RH”) is increased from about 0% to about 90% RH,Form C exhibits about 0.6% w/w water uptake. In certain embodiments,Form C comprises less than 0.1% water as determined in a coulometricKarl Fischer (KF) titrator equipped with an oven sample processor set at225° C.

In certain embodiments, Form C shows no significant degradation orresidual solvent by ¹H NMR (FIG. 21).

In certain embodiments, Form C of Compound 1 is characterized by itsstability profile upon compression. In certain embodiments, Form C isstable, e.g., its XRPD pattern remains substantially unchanged withbroader diffraction peaks, upon application of 2000-psi pressure forabout 1 minute (FIG. 22).

In still another embodiment, Form C of Compound 1 is substantially pure.In certain embodiments, the substantially pure Form C of Compound 1 issubstantially free of other solid forms, e.g., amorphous form. Incertain embodiments, the purity of the substantially pure Form C ofCompound 1 is no less than about 95% pure, no less than about 96% pure,no less than about 97% pure, no less than about 98% pure, no less thanabout 98.5% pure, no less than about 99% pure, no less than about 99.5%pure, or no less than about 99.8% pure.

Certain embodiments herein provide Form C of Compound 1 which issubstantially pure. Certain embodiments herein provide Form C ofCompound 1 which is substantially free of other solid forms comprisingCompound 1 including, e.g., Forms A, B, D, E, and/or an amorphous solidform comprising Compound 1 as provided herein. Certain embodimentsherein provide Form C as a mixture of solid forms comprising Compound 1,including, e.g., a mixture comprising one or more of the following:Forms A, B, D, E, and an amorphous solid form comprising Compound 1 asprovided herein.

6.2.4. Form D of Compound 1

In certain embodiments, provided herein is Form D of Compound 1. Incertain embodiments, Form D of Compound 1 is a DMSO solvate.

In certain embodiments, Form D is obtained by heating Form B inDMSO/methyl isobutyl ketone and cooling the solution.

In certain embodiments, Form D is crystalline, as indicated by, e.g.,X-ray powder diffraction measurements. In one embodiment, Form D ofCompound 1 has an X-ray powder diffraction pattern substantially asshown in FIG. 23.

In one embodiment, Form D of Compound 1 has one or more characteristicX-ray powder diffraction peaks at a two-theta angle of approximately14.1, 14.3, 18.8, 19.1, 23.6 or 24.0 degrees 2θ as depicted in FIG. 23.In another embodiment, Form D of Compound 1 has one, two, three or fourcharacteristic X-ray powder diffraction peaks at a two-theta angle ofapproximately 14.1, 14.3, 18.8 or 19.1 degrees 2θ. In anotherembodiment, Form D of Compound 1 has one, two, three, four, five, six orseven characteristic X-ray powder diffraction peaks as set forth inTable 4. In another embodiment, Form D of Compound 1 has one, two, orthree characteristic X-ray powder diffraction peaks as set forth inTable 4.

In one embodiment, provided herein is a crystalline form of Compound 1having a thermogravimetric (TGA) thermograph corresponding substantiallyto the representative TGA thermogram as depicted in FIG. 24. In certainembodiments, Form D shows TGA weight loss of about 14.1% up to 140° C.

In certain embodiments, Form D comprises DMSO in about 14.3 wt % asmeasured by gas chromatography.

In still another embodiment, Form D of Compound 1 is substantially pure.In certain embodiments, the substantially pure Form D of Compound 1 issubstantially free of other solid forms, e.g., amorphous form. Incertain embodiments, the purity of the substantially pure Form D ofCompound 1 is no less than about 95% pure, no less than about 96% pure,no less than about 97% pure, no less than about 98% pure, no less thanabout 98.5% pure, no less than about 99% pure, no less than about 99.5%pure, or no less than about 99.8% pure.

Certain embodiments herein provide Form D of Compound 1 which issubstantially pure. Certain embodiments herein provide Form D ofCompound 1 which is substantially free of other solid forms comprisingCompound 1 including, e.g., Forms A, B, C, E, and/or an amorphous solidform comprising Compound 1 as provided herein. Certain embodimentsherein provide Form D as a mixture of solid forms comprising Compound 1,including, e.g., a mixture comprising one or more of the following:Forms A, B, C, E, and an amorphous solid form comprising Compound 1 asprovided herein.

6.2.5. Form E of Compound 1

In certain embodiments, provided herein is Form E of Compound 1. Incertain embodiments, Form E of Compound 1 is a DMSO solvate.

In certain embodiments, Form E is obtained from Form C in DMSO/MIBK orDMSO/IPA or DMSO/anisole at room temperature.

In certain embodiments, Form E is crystalline, as indicated by, e.g.,X-ray powder diffraction measurements. In one embodiment, Form E ofCompound 1 has an X-ray powder diffraction pattern substantially asshown in FIG. 25.

In one embodiment, Form E of Compound 1 has one or more characteristicX-ray powder diffraction peaks at a two-theta angle of approximately10.5, 12.5, 16.1, 17.0, 18.5, 21.2, 21.7, 22.6, 22.9, 23.4, 23.8, 24.1,25.1 or 26.7, degrees 2θ as depicted in FIG. 25. In another embodiment,Form E of Compound 1 has one, two, three or four characteristic X-raypowder diffraction peaks at a two-theta angle of approximately 16.1,17.0, 21.2 or 22.9 degrees 2θ. In another embodiment, Form E of Compound1 has one, two, three, four, five, six or seven characteristic X-raypowder diffraction peaks as set forth in Table 5. In another embodiment,Form E of Compound 1 has one, two, or three characteristic X-ray powderdiffraction peaks as set forth in Table 5.

In one embodiment, provided herein is a crystalline form of Compound 1having a thermogravimetric (TGA) thermograph corresponding substantiallyto the representative TGA thermogram as depicted in FIG. 26. In certainembodiments, Form E shows TGA weight loss of about 19.4% up to 120° C.In certain embodiments, Form E shows additional weight loss of 24.9%between 120 and 220° C.

In one embodiment, Form E of Compound 1 is substantially pure. Incertain embodiments, the substantially pure Form E of Compound 1 issubstantially free of other solid forms, e.g., amorphous form. Incertain embodiments, the purity of the substantially pure Form E ofCompound 1 is no less than about 95% pure, no less than about 96% pure,no less than about 97% pure, no less than about 98% pure, no less thanabout 98.5% pure, no less than about 99% pure, no less than about 99.5%pure, or no less than about 99.8% pure.

Certain embodiments herein provide Form E of Compound 1 which issubstantially pure. Certain embodiments herein provide Form E ofCompound 1 which is substantially free of other solid forms comprisingCompound 1 including, e.g., Forms A, B, C, D and/or an amorphous solidform comprising Compound 1 as provided herein. Certain embodimentsherein provide Form C as a mixture of solid forms comprising Compound 1,including, e.g., a mixture comprising one or more of the following:Forms A, B, C, D and an amorphous solid form comprising Compound 1 asprovided herein.

6.2.6. Amorphous Compound 1

In certain embodiments, provided herein is amorphous Compound 1.

In certain embodiments, provided herein are methods for making theamorphous form by heating Compound 1 in THF and water, and lyophilizingthe solution. In one embodiment the lyophilizing comprises freezing thesolution and removing the solvent under reduced pressure.

In one embodiment, provided herein is an amorphous solid form ofCompound 1 having a DSC thermogram as depicted in FIG. 27.

In one embodiment, amorphous Compound 1 has an X-ray powder diffractionpattern substantially as shown in FIG. 28.

In one embodiment, amorphous Compound 1 has a ¹H NMR spectrumsubstantially as shown in FIG. 29.

In still another embodiment, amorphous Compound 1 is substantially pure.In certain embodiments, the substantially pure amorphous Compound 1 issubstantially free of other solid forms, e.g., Form A, Form B, Form C,Form D or Form E. In certain embodiments, the purity of thesubstantially pure amorphous Compound 1 is no less than about 95% pure,no less than about 96% pure, no less than about 97% pure, no less thanabout 98% pure, no less than about 98.5% pure, no less than about 99%pure, no less than about 99.5% pure, or no less than about 99.8% pure.

6.2.7. Isotopologues of Compound 1

Also provided herein are isotopically enriched analogs of the compounds(“isotopologues”) provided herein. Isotopic enrichment (for example,deuteration) of pharmaceuticals to improve pharmacokinetics (“PK”),pharmacodynamics (“PD”), and toxicity profiles, has been demonstratedpreviously with some classes of drugs. See, for example, Lijinsky et.al., Food Cosmet. Toxicol., 20: 393 (1982); Lijinsky et. al., J. Nat.Cancer Inst., 69: 1127 (1982); Mangold et. al., Mutation Res. 308: 33(1994); Gordon et. al., Drug Metab. Dispos., 15: 589 (1987); Zello et.al., Metabolism, 43: 487 (1994); Gately et. al., J. Nucl. Med., 27: 388(1986); Wade D, Chem. Biol. Interact. 117: 191 (1999).

Without being limited by any particular theory, iotopic enrichment of adrug can be used, for example, to (1) reduce or eliminate unwantedmetabolites, (2) increase the half-life of the parent drug, (3) decreasethe number of doses needed to achieve a desired effect, (4) decrease theamount of a dose necessary to achieve a desired effect, (5) increase theformation of active metabolites, if any are formed, and/or (6) decreasethe production of deleterious metabolites in specific tissues and/orcreate a more effective drug and/or a safer drug for combinationtherapy, whether the combination therapy is intentional or not.

Replacement of an atom for one of its isotopes often will result in achange in the reaction rate of a chemical reaction. This phenomenon isknown as the Kinetic Isotope Effect (“KIE”). For example, if a C—H bondis broken during a rate-determining step in a chemical reaction (i.e.the step with the highest transition state energy), substitution of adeuterium for that hydrogen will cause a decrease in the reaction rateand the process will slow down. This phenomenon is known as theDeuterium Kinetic Isotope Effect (“DKIE”). (See, e.g, Foster et al.,Adv. Drug Res., vol. 14, pp. 1-36 (1985); Kushner et al., Can. J.Physiol. Pharmacol., vol. 77, pp. 79-88 (1999)).

The magnitude of the DKIE can be expressed as the ratio between therates of a given reaction in which a C—H bond is broken, and the samereaction where deuterium is substituted for hydrogen. The DKIE can rangefrom about 1 (no isotope effect) to very large numbers, such as 50 ormore, meaning that the reaction can be fifty, or more, times slower whendeuterium is substituted for hydrogen. Without being limited by aparticular theory, high DKIE values may be due in part to a phenomenonknown as tunneling, which is a consequence of the uncertainty principle.Tunneling is ascribed to the small mass of a hydrogen atom, and occursbecause transition states involving a proton can sometimes form in theabsence of the required activation energy. Because deuterium has moremass than hydrogen, it statistically has a much lower probability ofundergoing this phenomenon.

Tritium (“T”) is a radioactive isotope of hydrogen, used in research,fusion reactors, neutron generators and radiopharmaceuticals. Tritium isa hydrogen atom that has 2 neutrons in the nucleus and has an atomicweight close to 3. It occurs naturally in the environment in very lowconcentrations, most commonly found as T₂O. Tritium decays slowly(half-life=12.3 years) and emits a low energy beta particle that cannotpenetrate the outer layer of human skin. Internal exposure is the mainhazard associated with this isotope, yet it must be ingested in largeamounts to pose a significant health risk. As compared with deuterium, alesser amount of tritium must be consumed before it reaches a hazardouslevel. Substitution of tritium (“T”) for hydrogen results in yet astronger bond than deuterium and gives numerically larger isotopeeffects.

Similarly, substitution of isotopes for other elements, including, butnot limited to, ¹³C or ¹⁴C for carbon, ³³S, ³⁴S, or ³⁶S for sulfur, ¹⁵Nfor nitrogen, and ¹⁷O or ¹⁸O for oxygen, will provide a similar kineticisotope effects.

6.3. Methods of Treatment

In one embodiment, provided herein is a method of treating andpreventing cancer, which comprises administering to a patient a solidform of Compound 1 provided herein.

In another embodiment, provided herein is method of managing cancer,which comprises administering to a patient a solid form of Compound 1provided herein.

Also provided herein are methods of treating patients who have beenpreviously treated for cancer but are non-responsive to standardtherapies, as well as those who have not previously been treated. Theinvention also encompasses methods of treating patients regardless ofpatient's age, although some diseases or disorders are more common incertain age groups. The invention further encompasses methods oftreating patients who have undergone surgery in an attempt to treat thedisease or condition at issue, as well as those who have not. Becausepatients with cancer have heterogeneous clinical manifestations andvarying clinical outcomes, the treatment given to a patient may vary,depending on his/her prognosis. The skilled clinician will be able toreadily determine without undue experimentation specific secondaryagents, types of surgery, and types of non-drug based standard therapythat can be effectively used to treat an individual patient with cancer.

As used herein, the term “cancer” includes, but is not limited to, solidtumors and blood borne tumors. The term “cancer” refers to disease ofskin tissues, organs, blood, and vessels, including, but not limited to,cancers of the bladder, bone, blood, brain, breast, cervix, chest,colon, endrometrium, esophagus, eye, head, kidney, liver, lymph nodes,lung, mouth, neck, ovaries, pancreas, prostate, rectum, stomach, testis,throat, and uterus. Specific cancers include, but are not limited to,advanced malignancy, amyloidosis, neuroblastoma, meningioma,hemangiopericytoma, multiple brain metastase, glioblastoma multiforms,glioblastoma, brain stem glioma, poor prognosis malignant brain tumor,malignant glioma, recurrent malignant giolma, anaplastic astrocytoma,anaplastic oligodendroglioma, neuroendocrine tumor, rectaladenocarcinoma, colorectal cancer, including stage 3 and stage 4colorectal cancer, unresectable colorectal carcinoma, metastatichepatocellular carcinoma, Kaposi's sarcoma, karotype acute myeloblasticleukemia, Hodgkin's lymphoma, non-Hodgkin's lymphoma, cutaneous T-Celllymphoma, cutaneous B-Cell lymphoma, diffuse large B-Cell lymphoma, lowgrade follicular lymphoma, malignant melanoma, malignant mesothelioma,malignant pleural effusion mesothelioma syndrome, peritoneal carcinoma,papillary serous carcinoma, gynecologic sarcoma, soft tissue sarcoma,scleroderma, cutaneous vasculitis, Langerhans cell histiocytosis,leiomyosarcoma, fibrodysplasia ossificans progressive, hormonerefractory prostate cancer, resected high-risk soft tissue sarcoma,unrescectable hepatocellular carcinoma, Waldenstrom's macroglobulinemia,smoldering myeloma, indolent myeloma, fallopian tube cancer, androgenindependent prostate cancer, androgen dependent stage IV non-metastaticprostate cancer, hormone-insensitive prostate cancer,chemotherapy-insensitive prostate cancer, papillary thyroid carcinoma,follicular thyroid carcinoma, medullary thyroid carcinoma, andleiomyoma.

In certain embodiments, the cancer is a solid tumor. In certainembodiments, the solid tumor is metastatic. In certain embodiments, thesolid tumor is drug-resistant. In certain embodiments, the solid tumoris hepatocellular carcinoma, prostate cancer, ovarian cancer, orglioblastoma.

In certain embodiments, the cancer is a blood borne tumor. In certainembodiments, the blood borne tumor is metastatic. In certainembodiments, the blood borne tumor is drug resistant. In certainembodiments, the cancer is leukemia.

In one embodiment, methods provided herein encompass treating,preventing and/or managing various types of leukemias such as chroniclymphocytic leukemia (CLL), chronic myelocytic leukemia (CML), acutelymphoblastic leukemia (ALL), acute myeloid leukemia (AML), and acutemyeloblastic leukemia (AML) by administering a therapeutically effectiveamount of a solid form of Compound 1 provided herein.

In some embodiments, the methods provided herein encompass treating,preventing and/or managing acute leukemia in a subject. In someembodiments, the acute leukemia is acute myeloid leukemia (AML), whichincludes, but is not limited to, undifferentiated AML (M0), myeloblasticleukemia (M1), myeloblastic leukemia (M2), promyelocytic leukemia (M3 orM3 variant (M3V)), myelomonocytic leukemia (M4 or M4 variant witheosinophilia (M4E)), monocytic leukemia (M5), erythroleukemia (M6), andmegakaryoblastic leukemia (M7). In one embodiment, the acute myeloidleukemia is undifferentiated AML (M0). In one embodiment, the acutemyeloid leukemia is myeloblastic leukemia (M1). In one embodiment, theacute myeloid leukemia is myeloblastic leukemia (M2). In one embodiment,the acute myeloid leukemia is promyelocytic leukemia (M3 or M3 variant(M3V)). In one embodiment, the acute myeloid leukemia is myelomonocyticleukemia (M4 or M4 variant with eosinophilia (M4E)). In one embodiment,the acute myeloid leukemia is monocytic leukemia (M5). In oneembodiment, the acute myeloid leukemia is erythroleukemia (M6). In oneembodiment, the acute myeloid leukemia is megakaryoblastic leukemia(M7).

In certain embodiments, the methods of treating, preventing and/ormanaging acute myeloid leukemia in a subject comprise the step ofadministering to the subject an amount of a solid form of Compound 1provided herein effective to treat, prevent and/or manage acute myeloidleukemia alone or in combination.

In one embodiment, provided herein are methods of treating, preventingand/or managing acute myeloid leukemia by intravenous administration ofa solid form of Compound 1. In one embodiment, the solid form ofCompound 1 is dissolved in water to form an aqueous solution forintravenous administration in methods of treating, preventing and/ormanaging acute myeloid leukemia provided herein.

In one embodiment, provided herein are methods of treating, preventingand/or managing acute myeloid leukemia by intravenous administration ofForm C of Compound 1. In one embodiment, Form C of Compound 1 isdissolved in water to form an aqueous solution for intravenousadministration in methods of treating, preventing and/or managing acutemyeloid leukemia provided herein.

In some embodiments, the methods comprise the step of administering tothe subject a solid form of Compound 1 provided herein in combinationwith a second active agent in amounts effective to treat, prevent and/ormanage acute myeloid leukemia.

In some embodiments, the methods provided herein encompass treating,preventing and/or managing acute lymphocytic leukemia (ALL) in asubject. In some embodiments, acute lymphocytic leukemia includesleukemia that originates in the blast cells of the bone marrow(B-cells), thymus (T-cells), and lymph nodes. The acute lymphocyticleukemia can be categorized according to the French-American-British(FAB) Morphological Classification Scheme as L1—Mature-appearinglymphoblasts (T-cells or pre-B-cells), L2—Immature and pleomorphic(variously shaped) lymphoblasts (T-cells or pre-B-cells), andL3—Lymphoblasts (B-cells; Burkitt's cells). In one embodiment, the acutelymphocytic leukemia originates in the blast cells of the bone marrow(B-cells). In one embodiment, the acute lymphocytic leukemia originatesin the thymus (T-cells). In one embodiment, the acute lymphocyticleukemia originates in the lymph nodes. In one embodiment, the acutelymphocytic leukemia is L1 type characterized by mature-appearinglymphoblasts (T-cells or pre-B-cells). In one embodiment, the acutelymphocytic leukemia is L2 type characterized by immature andpleomorphic (variously shaped) lymphoblasts (T-cells or pre-B-cells). Inone embodiment, the acute lymphocytic leukemia is L3 type characterizedby lymphoblasts (B-cells; Burkitt's cells). In certain embodiments, theacute lymphocytic leukemia is T-cell leukemia. In one embodiment, theT-cell leukemia is peripheral T-cell leukemia. In another embodiment,the T-cell leukemia is T-cell lymphoblastic leukemia. In anotherembodiment, the T-cell leukemia is cutaneous T-cell leukemia. In anotherembodiment, the T-cell leukemia is adult T-cell leukemia. Thus, themethods of treating, preventing and/or managing acute lymphocyticleukemia in a subject comprise the step of administering to the subjectan amount of a solid form of Compound 1 provided herein effective totreat, prevent and/or manage acute lymphocytic leukemia alone or incombination with a second active agent. In some embodiments, the methodscomprise the step of administering to the subject a solid form ofCompound 1 provided herein in combination with a second active agent inamounts effective to treat, prevent and/or manage acute lymphocyticleukemia.

In some embodiments, the methods provided herein encompass treating,preventing and/or managing chronic myelogenous leukemia (CML) in asubject. The methods comprise the step of administering to the subjectan amount of a solid form of Compound 1 provided herein effective totreat, prevent and/or manage chronic myelogenous leukemia. In someembodiments, the methods comprise the step of administering to thesubject a solid form of Compound 1 provided herein in combination with asecond active agent in amounts effective to treat, prevent and/or managechronic myelogenous leukemia.

In some embodiments, the methods provided herein encompass treating,preventing and/or managing chronic lymphocytic leukemia (CLL) in asubject. The methods comprise the step of administering to the subjectan amount of a solid form of Compound 1 provided herein effective totreat, prevent and/or manage chronic lymphocytic leukemia. In someembodiments, the methods comprise the step of administering to thesubject a solid form of Compound 1 provided herein in combination with asecond active agent in amounts effective to treat, prevent and/or managechronic lymphocytic leukemia.

In certain embodiments, provided herein are methods of treating,preventing, and/or managing disease in patients with impaired renalfunction. In certain embodiments, provided herein are method oftreating, preventing, and/or managing cancer in patients with impairedrenal function. In certain embodiments, provided herein are methods ofproviding appropriate dose adjustments for patients with impaired renalfunction due to, but not limited to, disease, aging, or other patientfactors.

In certain embodiments, provided herein are methods of treating,preventing, and/or managing lymphoma, including non-Hodgkin's lymphoma.In some embodiments, provided herein are methods for the treatmentand/or management of non-Hodgkin's lymphoma (NHL), including but notlimited to, diffuse large B-cell lymphoma (DLBCL), using prognosticfactors.

In certain embodiments, provided herein are methods of treating,preventing, and/or managing multiple myeloma, includingrelapsed/refractory multiple myeloma in patients with impaired renalfunction or a symptom thereof, comprising administering atherapeutically effective amount of a solid form of Compound 1 providedherein to a patient having relapsed/refractory multiple myeloma withimpaired renal function.

In one embodiment, provided herein are methods of treating, preventing,and/or managing a myelodysplastic syndrome (MDS) by administering atherapeutically active amount of a solid form of Compound 1 providedherein. In one embodiment, the MDS is relapsed, resistant or refractoryMDS. In one embodiment, MDS is selected from refractory anemia (RA); RAwith ringed sideroblasts (RARS); RA with excess of blasts (RAEB);refractory cytopenia with multilineage dysplasia (RCMD), refractorycytopenia with unilineage dysplasia (RCUD); unclassifiablemyelodysplastic syndrome (MDS-U), myelodysplastic syndrome associatedwith an isolated del(5q) chromosome abnormality, therapy-related myeloidneoplasms and chronic myelomonocytic leukemia (CMML).

In certain embodiments, a therapeutically or prophylactically effectiveamount of a solid form of Compound 1 provided herein is from about 0.005to about 1,000 mg per day, from about 0.01 to about 500 mg per day, fromabout 0.01 to about 250 mg per day, from about 0.01 to about 100 mg perday, from about 0.1 to about 100 mg per day, from about 0.5 to about 100mg per day, from about 1 to about 100 mg per day, from about 0.01 toabout 50 mg per day, from about 0.1 to about 50 mg per day, from about0.5 to about 50 mg per day, from about 1 to about 50 mg per day, fromabout 0.02 to about 25 mg per day, from about 0.05 to about 10 mg perday, from about 0.05 to about 5 mg per day, from about 0.1 to about 5 mgper day, or from about 0.5 to about 5 mg per day.

In certain embodiments, the therapeutically or prophylacticallyeffective amount is about 0.1, about 0.2, about 0.5, about 1, about 2,about 3, about 4, about 5, about 6, about 7, about 8, about 9, about 10,about 15, about 20, about 25, about 30, about 40, about 45, about 50,about 60, about 70, about 80, about 90, about 100, or about 150 mg perday. In some such embodiments, the therapeutically or prophylacticallyeffective amount is about 2, about 3, about 4, about 5, about 6 or about7 mg per day.

In one embodiment, the recommended daily dose range of a solid form ofCompound 1 provided herein, for the conditions described herein liewithin the range of from about 0.05 mg to about 50 mg per day,preferably given as a single once-a-day dose, or in divided dosesthroughout a day. In some embodiments, the dosage ranges from about 1 mgto about 50 mg per day. In other embodiments, the dosage ranges fromabout 0.5 to about 5 mg per day. Specific doses per day include 0.1,0.2, 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18,19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36,37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49 or 50 mg per day.

In a specific embodiment, the recommended starting dosage may be 0.1,0.5, 1, 2, 3, 4, 5, 10, 15, 20, 25 or 50 mg per day. In anotherembodiment, the recommended starting dosage may be 0.1, 0.5, 1, 2, 3, 4,or 5 mg per day. The dose may be escalated to 15, 20, 25, 30, 35, 40, 45and 50 mg/day. In a specific embodiment, a solid form of Compound 1 canbe administered in an amount of about 25 mg/day to patients withleukemia, including AML. In a particular embodiment, a solid form ofCompound 1 can be administered in an amount of about 10 mg/day topatients with leukemia, including AML. In a particular embodiment, asolid form of Compound 1 can be administered in an amount of about 5mg/day to patients with leukemia, including AML. In a particularembodiment, a solid form of Compound 1 can be administered in an amountof about 4 mg/day to patients with leukemia, including AML. In aparticular embodiment, a solid form of Compound 1 provided herein can beadministered in an amount of about 3 mg/day to patients with leukemia,including AML. In a particular embodiment, Compound 1 provided hereincan be administered in an amount of about 2 mg/day to patients withleukemia, including AML. In a particular embodiment, Compound 1 providedherein can be administered in an amount of about 1 mg/day to patientswith leukemia, including AML. In a particular embodiment, Compound 1provided herein can be administered in an amount of about 0.5 mg/day topatients with leukemia, including AML.

In a specific embodiment, a solid form of Compound 1 can be administeredin an amount of about 25 mg/day to patients with MDS. In a particularembodiment, a solid form of Compound 1 can be administered in an amountof about 10 mg/day to patients with MDS. In a particular embodiment, asolid form of Compound 1 can be administered in an amount of about 5mg/day to patients with MDS. In a particular embodiment, a solid form ofCompound 1 can be administered in an amount of about 4 mg/day topatients with MDS. In a particular embodiment, a solid form of Compound1 provided herein can be administered in an amount of about 3 mg/day topatients with MDS. In a particular embodiment, a solid form of Compound1 provided herein can be administered in an amount of about 2 mg/day topatients with MDS. In a particular embodiment, a solid form of Compound1 provided herein can be administered in an amount of about 1 mg/day topatients with MDS. In a particular embodiment, a solid form of Compound1 provided herein can be administered in an amount of about 0.5 mg/dayto patients with MDS.

In certain embodiments, the therapeutically or prophylacticallyeffective amount is from about 0.001 to about 100 mg/kg/day, from about0.01 to about 50 mg/kg/day, from about 0.01 to about 25 mg/kg/day, fromabout 0.01 to about 10 mg/kg/day, from about 0.01 to about 9 mg/kg/day,0.01 to about 8 mg/kg/day, from about 0.01 to about 7 mg/kg/day, fromabout 0.01 to about 6 mg/kg/day, from about 0.01 to about 5 mg/kg/day,from about 0.01 to about 4 mg/kg/day, from about 0.01 to about 3mg/kg/day, from about 0.01 to about 2 mg/kg/day, from about 0.01 toabout 1 mg/kg/day, or from about 0.01 to about 0.05 mg/kg/day.

The administered dose can also be expressed in units other thanmg/kg/day. For example, doses for parenteral administration can beexpressed as mg/m²/day. One of ordinary skill in the art would readilyknow how to convert doses from mg/kg/day to mg/m²/day to given eitherthe height or weight of a subject or both (see,www.fda.gov/cder/cancer/animalframe.htm). For example, a dose of 1mg/kg/day for a 65 kg human is approximately equal to 38 mg/m²/day.

In certain embodiments, the amount of a solid form of Compound 1administered is sufficient to provide a plasma concentration of thecompound at steady state, ranging from about 0.001 to about 500 μM,about 0.002 to about 200 μM, about 0.005 to about 100 μM, about 0.01 toabout 50 μM, from about 1 to about 50 μM, about 0.02 to about 25 μM,from about 0.05 to about 20 μM, from about 0.1 to about 20 μM, fromabout 0.5 to about 20 μM, or from about 1 to about 20 μM.

In other embodiments, the amount of a solid form of Compound 1administered is sufficient to provide a plasma concentration of thecompound at steady state, ranging from about 5 to about 100 nM, about 5to about 50 nM, about 10 to about 100 nM, about 10 to about 50 nM orfrom about 50 to about 100 nM.

As used herein, the term “plasma concentration at steady state” is theconcentration reached after a period of administration of a solid formof Compound 1 provided herein. Once steady state is reached, there areminor peaks and troughs on the time dependent curve of the plasmaconcentration of the solid form.

In certain embodiments, the amount of a solid form of Compound 1administered is sufficient to provide a maximum plasma concentration(peak concentration) of the compound, ranging from about 0.001 to about500 μM, about 0.002 to about 200 μM, about 0.005 to about 100 μM, about0.01 to about 50 μM, from about 1 to about 50 μM, about 0.02 to about 25μM, from about 0.05 to about 20 μM, from about 0.1 to about 20 μM, fromabout 0.5 to about 20 μM, or from about 1 to about 20 μM.

In certain embodiments, the amount of a solid form of Compound 1administered is sufficient to provide a minimum plasma concentration(trough concentration) of the compound, ranging from about 0.001 toabout 500 μM, about 0.002 to about 200 μM, about 0.005 to about 100 μM,about 0.01 to about 50 μM, from about 1 to about 50 μM, about 0.01 toabout 25 μM, from about 0.01 to about 20 μM, from about 0.02 to about 20μM, from about 0.02 to about 20 μM, or from about 0.01 to about 20 μM.

In certain embodiments, the amount of a solid form of Compound 1administered is sufficient to provide an area under the curve (AUC) ofthe compound, ranging from about 100 to about 100,000 ng*hr/mL, fromabout 1,000 to about 50,000 ng*hr/mL, from about 5,000 to about 25,000ng*hr/mL, or from about 5,000 to about 10,000 ng*hr/mL.

In certain embodiments, the patient to be treated with one of themethods provided herein has not been treated with anticancer therapyprior to the administration of a solid form of Compound 1 providedherein. In certain embodiments, the patient to be treated with one ofthe methods provided herein has been treated with anticancer therapyprior to the administration of a solid form of Compound 1 providedherein. In certain embodiments, the patient to be treated with one ofthe methods provided herein has developed drug resistance to theanticancer therapy.

The methods provided herein encompass treating a patient regardless ofpatient's age, although some diseases or disorders are more common incertain age groups.

Depending on the disease to be treated and the subject's condition, asolid form of Compound 1 provided herein may be administered byparenteral (e.g., intramuscular, intraperitoneal, intravenous, CIV,intracistemal injection or infusion, subcutaneous injection, orimplant), inhalation, nasal, vaginal, rectal, sublingual, or topical(e.g., transdermal or local) routes of administration. The solid formsof Compound 1 may be formulated, alone or together, in suitable dosageunit with pharmaceutically acceptable excipients, carriers, adjuvantsand vehicles, appropriate for each route of administration.

In another embodiment, the solid form of Compound 1 is administeredparenterally. In certain embodiments, an aqueous solution containing oneor more solid forms of Compound 1 is administered parenterally.

In yet another embodiment, a solid form of Compound 1 provided herein isadministered intravenously. In certain embodiments, an aqueous solutioncontaining one or more solid forms of Compound 1 is administeredintravenously.

The solid forms of Compound 1 provided herein can be delivered as asingle dose such as, e.g., a single bolus injection, or over time, suchas, e.g., continuous infusion over time or divided bolus doses overtime. The solid forms of Compound 1 can be administered repeatedly ifnecessary, for example, until the patient experiences stable disease orregression, or until the patient experiences disease progression orunacceptable toxicity. For example, stable disease for solid tumorsgenerally means that the perpendicular diameter of measurable lesionshas not increased by 25% or more from the last measurement. ResponseEvaluation Criteria in Solid Tumors (RECIST) Guidelines, Journal of theNational Cancer Institute 92(3): 205-216 (2000). Stable disease or lackthereof is determined by methods known in the art such as evaluation ofpatient symptoms, physical examination, visualization of the tumor thathas been imaged using X-ray, CAT, PET, or MM scan and other commonlyaccepted evaluation modalities.

The solid forms of Compound 1 provided herein can be administered oncedaily (QD), or divided into multiple daily doses such as twice daily(BID), three times daily (TID), and four times daily (QID). In addition,the administration can be continuous (i.e., daily for consecutive daysor every day), intermittent, e.g., in cycles (i.e., including days,weeks, or months of rest without drug). As used herein, the term “daily”is intended to mean that a therapeutic compound, such as a solid form ofCompound 1, is administered once or more than once each day, forexample, for a period of time. The term “continuous” is intended to meanthat a therapeutic compound, such as a solid form of Compound 1, isadministered daily for an uninterrupted period of at least 10 days to 52weeks. The term “intermittent” or “intermittently” as used herein isintended to mean stopping and starting at either regular or irregularintervals. For example, intermittent administration of the solid form ofCompound 1 is administration for one to six days per week,administration in cycles (e.g., daily administration for two to eightconsecutive weeks, then a rest period with no administration for up toone week), or administration on alternate days. The term “cycling” asused herein is intended to mean that a therapeutic compound, such as asolid form of Compound 1, is administered daily or continuously but witha rest period. In some such embodiments, administration is once a dayfor two to six days, then a rest period with no administration for fiveto seven days. In some other such embodiments, administration is once aday for the first two to five or ten days of a 28 day cycle, followed bya rest period with no administration for the rest of the 28 day cycle.

In some embodiments, the frequency of administration is in the range ofabout a daily dose to about a monthly dose. In certain embodiments,administration is once a day, twice a day, three times a day, four timesa day, once every other day, twice a week, once every week, once everytwo weeks, once every three weeks, or once every four weeks. In oneembodiment, a solid form of Compound 1 is administered once a day. Inanother embodiment, a solid form of Compound 1 is administered twice aday. In yet another embodiment, a solid form of Compound 1 providedherein is administered three times a day. In still another embodiment, asolid form of Compound 1 provided herein is administered four times aday.

In certain embodiments, a solid form of Compound 1 provided herein isadministered once per day from one day to six months, from one week tothree months, from one week to four weeks, from one week to three weeks,or from one week to two weeks. In certain embodiments, a solid form ofCompound 1 provided herein is administered once per day for one week,two weeks, three weeks, or four weeks. In one embodiment, a solid formof Compound 1 provided herein is administered once per day for 4 days.In one embodiment, a solid form of Compound 1 provided herein isadministered once per day for 5 days. In one embodiment, a solid form ofCompound 1 provided herein is administered once per day for 6 days. Inone embodiment, a solid form of Compound 1 provided herein isadministered once per day for one week. In another embodiment, a solidform of Compound 1 provided herein is administered once per day for twoweeks. In yet another embodiment, a solid form of Compound 1 providedherein is administered once per day for three weeks. In still anotherembodiment, a solid form of Compound 1 provided herein is administeredonce per day for four weeks.

6.3.1. Combination Therapy With A Second Active Agent

The solid forms of Compound 1 provided herein can also be combined orused in combination with other therapeutic agents useful in thetreatment and/or prevention of cancer described herein.

In one embodiment, provided herein is a method of treating, preventing,and/or managing cancer, comprising administering to a patient a solidform of Compound 1 provided herein in combination with one or moresecond active agents, and optionally in combination with radiationtherapy, blood transfusions, or surgery. Examples of second activeagents are disclosed herein.

The solid forms of Compound 1 provided herein can also be combined orused in combination with other therapeutic agents useful in thetreatment and/or prevention of MDS described herein.

In one embodiment, provided herein is a method of treating, preventing,and/or managing MDS, comprising administering to a patient a solid formof Compound 1 provided herein in combination with one or more secondactive agents, and optionally in combination with radiation therapy,blood transfusions, or surgery. Examples of second active agents aredisclosed herein.

As used herein, the term “in combination” includes the use of more thanone therapy (e.g., one or more prophylactic and/or therapeutic agents).However, the use of the term “in combination” does not restrict theorder in which therapies (e.g., prophylactic and/or therapeutic agents)are administered to a patient with a disease or disorder. A firsttherapy (e.g., a prophylactic or therapeutic agent such as a compoundprovided herein, a compound provided herein, e.g., a solid form ofCompound 1 can be administered prior to (e.g., 5 minutes, 15 minutes, 30minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 24hours, 48 hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks,5 weeks, 6 weeks, 8 weeks, or 12 weeks before), concomitantly with, orsubsequent to (e.g., 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1hour, 2 hours, 4 hours, 6 hours, 12 hours, 24 hours, 48 hours, 72 hours,96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks,or 12 weeks after) the administration of a second therapy (e.g., aprophylactic or therapeutic agent) to the subject. Triple therapy isalso contemplated herein.

Administration of a solid form of Compound 1 provided herein and one ormore second active agents to a patient can occur simultaneously orsequentially by the same or different routes of administration. Thesuitability of a particular route of administration employed for aparticular active agent will depend on the active agent itself (e.g.,whether it can be administered orally without decomposing prior toentering the blood stream) and the cancer being treated.

The route of administration of a solid form of Compound 1 is independentof the route of administration of a second therapy. In anotherembodiment, solid form of Compound 1 is administered intravenously.Thus, in accordance with these embodiments, a solid form of Compound 1is administered intravenously, and the second therapy can beadministered orally, parenterally, intraperitoneally, intravenously,intraarterially, transdermally, sublingually, intramuscularly, rectally,transbuccally, intranasally, liposomally, via inhalation, vaginally,intraoccularly, via local delivery by catheter or stent, subcutaneously,intraadiposally, intraarticularly, intrathecally, or in a slow releasedosage form. In one embodiment, a solid form of Compound 1 and a secondtherapy are administered by the same mode of administration, orally orby IV. In another embodiment, a solid form of Compound 1 is administeredby one mode of administration, e.g., by IV, whereas the second agent (ananticancer agent) is administered by another mode of administration,e.g., orally.

In one embodiment, the second active agent is administered intravenouslyor subcutaneously and once or twice daily in an amount of from about 1to about 1000 mg, from about 5 to about 500 mg, from about 10 to about350 mg, or from about 50 to about 200 mg. The specific amount of thesecond active agent will depend on the specific agent used, the type ofdisease being treated and/or managed, the severity and stage of disease,and the amount of the solid form of Compound 1 provided herein and anyoptional additional active agents concurrently administered to thepatient.

One or more second active ingredients or agents can be used togetherwith a solid form of Compound 1 in the methods and compositions providedherein. Second active agents can be large molecules (e.g., proteins) orsmall molecules (e.g., synthetic inorganic, organometallic, or organicmolecules).

Examples of large molecule active agents include, but are not limitedto, hematopoietic growth factors, cytokines, and monoclonal andpolyclonal antibodies, particularly, therapeutic antibodies to cancerantigens. Typical large molecule active agents are biological molecules,such as naturally occurring or synthetic or recombinant proteins.Proteins that are particularly useful in the methods and compositionsprovided herein include proteins that stimulate the survival and/orproliferation of hematopoietic precursor cells and immunologicallyactive poietic cells in vitro or in vivo. Other useful proteinsstimulate the division and differentiation of committed erythroidprogenitors in cells in vitro or in vivo. Particular proteins include,but are not limited to: interleukins, such as IL-2 (includingrecombinant IL-II (“rIL2”) and canarypox IL-2), IL-10, IL-12, and IL-18;interferons, such as interferon alfa-2a, interferon alfa-2b, interferonalfa-n1, interferon alfa-n3, interferon beta-I a, and interferon gamma-Ib; GM-CF and GM-CSF; and EPO.

In certain embodiments, GM-CSF, G-CSF, SCF or EPO is administeredsubcutaneously during about five days in a four or six week cycle in anamount ranging from about 1 to about 750 mg/m²/day, from about 25 toabout 500 mg/m²/day, from about 50 to about 250 mg/m²/day, or from about50 to about 200 mg/m²/day. In certain embodiments, GM-CSF may beadministered in an amount of from about 60 to about 500 mcg/m²intravenously over 2 hours or from about 5 to about 12 mcg/m²/daysubcutaneously. In certain embodiments, G-CSF may be administeredsubcutaneously in an amount of about 1 mcg/kg/day initially and can beadjusted depending on rise of total granulocyte counts. The maintenancedose of G-CSF may be administered in an amount of about 300 (in smallerpatients) or 480 mcg subcutaneously. In certain embodiments, EPO may beadministered subcutaneously in an amount of 10,000 Unit 3 times perweek.

Particular proteins that can be used in the methods and compositionsinclude, but are not limited to: filgrastim, which is sold in the UnitedStates under the trade name Neupogen® (Amgen, Thousand Oaks, Calif.);sargramostim, which is sold in the United States under the trade nameLeukine® (Immunex, Seattle, Wash.); and recombinant EPO, which is soldin the United States under the trade name Epogen® (Amgen, Thousand Oaks,Calif.).

Recombinant and mutated forms of GM-CSF can be prepared as described inU.S. Pat. Nos. 5,391,485; 5,393,870; and 5,229,496; all of which areincorporated herein by reference. Recombinant and mutated forms of G-CSFcan be prepared as described in U.S. Pat. Nos. 4,810,643; 4,999,291;5,528,823; and 5,580,755; the entireties of which are incorporatedherein by reference.

Also provided for use in combination with a solid form of Compound 1provided herein are native, naturally occurring, and recombinantproteins. Further encompassed are mutants and derivatives (e.g.,modified forms) of naturally occurring proteins that exhibit, in vivo,at least some of the pharmacological activity of the proteins upon whichthey are based. Examples of mutants include, but are not limited to,proteins that have one or more amino acid residues that differ from thecorresponding residues in the naturally occurring forms of the proteins.Also encompassed by the term “mutants” are proteins that lackcarbohydrate moieties normally present in their naturally occurringforms (e.g., nonglycosylated forms). Examples of derivatives include,but are not limited to, pegylated derivatives and fusion proteins, suchas proteins formed by fusing IgG1 or IgG3 to the protein or activeportion of the protein of interest. See, e.g., Penichet, M. L. andMorrison, S. L., J. Immunol. Methods 248:91-101 (2001).

Antibodies that can be used in combination with a solid form of Compound1 provided herein include monoclonal and polyclonal antibodies. Examplesof antibodies include, but are not limited to, trastuzumab (Herceptin®),rituximab (Rituxan®), bevacizumab (Avastin™), pertuzumab (Omnitarg™),tositumomab (Bexxar®), edrecolomab (Panorex®), and G250. The solid formsof Compound 1 can also be combined with, or used in combination with,anti-TNF-α antibodies, and/or anti-EGFR antibodies, such as, forexample, Erbitux® or panitumumab.

Large molecule active agents may be administered in the form ofanti-cancer vaccines. For example, vaccines that secrete, or cause thesecretion of, cytokines such as IL-2, G-CSF, and GM-CSF can be used inthe methods and pharmaceutical compositions provided. See, e.g., Emens,L. A., et al., Curr. Opinion Mol. Ther. 3(1):77-84 (2001).

Second active agents that are small molecules can also be used toalleviate adverse effects associated with the administration of a solidform of Compound 1 provided herein. However, like some large molecules,many are believed to be capable of providing a synergistic effect whenadministered with (e.g., before, after or simultaneously) a solid formof Compound 1 provided herein. Examples of small molecule second activeagents include, but are not limited to, anti-cancer agents, antibiotics,immunosuppressive agents, and steroids.

In certain embodiments, the second agent is an HSP inhibitor, aproteasome inhibitor, a FLT3 inhibitior or a TOR kinase inhibitor.

Examples of anti-cancer agents to be used within the methods orcompositions described herein include, but are not limited to: acivicin;aclarubicin; acodazole hydrochloride; acronine; adozelesin; aldesleukin;altretamine; ambomycin; ametantrone acetate; amsacrine; anastrozole;anthramycin; asparaginase; asperlin; azacitidine; azetepa; azotomycin;batimastat; benzodepa; bicalutamide; bisantrene hydrochloride; bisnafidedimesylate; bizelesin; bleomycin sulfate; brequinar sodium; bropirimine;busulfan; cactinomycin; calusterone; caracemide; carbetimer;carboplatin; carmustine; carubicin hydrochloride; carzelesin;cedefingol; celecoxib (COX-2 inhibitor); chlorambucil; cirolemycin;cisplatin; cladribine; clofarabine; crisnatol mesylate;cyclophosphamide; Ara-C; dacarbazine; dactinomycin; daunorubicinhydrochloride; decitabine; dexormaplatin; dezaguanine; dezaguaninemesylate; diaziquone; docetaxel; doxorubicin; doxorubicin hydrochloride;droloxifene; droloxifene citrate; dromostanolone propionate; duazomycin;edatrexate; eflornithine hydrochloride; elsamitrucin; enloplatin;enpromate; epipropidine; epirubicin hydrochloride; erbulozole;esorubicin hydrochloride; estramustine; estramustine phosphate sodium;etanidazole; etoposide; etoposide phosphate; etoprine; fadrozolehydrochloride; fazarabine; fenretinide; floxuridine; fludarabinephosphate; fluorouracil; flurocitabine; fosquidone; fostriecin sodium;gemcitabine; gemcitabine hydrochloride; hydroxyurea; idarubicinhydrochloride; ifosfamide; ilmofosine; iproplatin; irinotecan;irinotecan hydrochloride; lanreotide acetate; letrozole; leuprolideacetate; liarozole hydrochloride; lometrexol sodium; lomustine;losoxantrone hydrochloride; masoprocol; maytansine; mechlorethaminehydrochloride; megestrol acetate; melengestrol acetate; melphalan;menogaril; mercaptopurine; methotrexate; methotrexate sodium; metoprine;meturedepa; mitindomide; mitocarcin; mitocromin; mitogillin; mitomalcin;mitomycin; mitosper; mitotane; mitoxantrone hydrochloride; mycophenolicacid; nocodazole; nogalamycin; omacetaxine; ormaplatin; oxisuran;paclitaxel; pegaspargase; peliomycin; pentamustine; peplomycin sulfate;perfosfamide; pipobroman; piposulfan; piroxantrone hydrochloride;plicamycin; plomestane; porfimer sodium; porfiromycin; prednimustine;procarbazine hydrochloride; puromycin; puromycin hydrochloride;pyrazofurin; riboprine; safingol; safingol hydrochloride; semustine;simtrazene; sorafenib; sparfosate sodium; sparsomycin; spirogermaniumhydrochloride; spiromustine; spiroplatin; streptonigrin; streptozocin;sulofenur; talisomycin; tecogalan sodium; taxotere; tegafur;teloxantrone hydrochloride; temoporfin; teniposide; teroxirone;testolactone; thiamiprine; thioguanine; thiotepa; tiazofurin;tirapazamine; toremifene citrate; trestolone acetate; triciribinephosphate; trimetrexate; trimetrexate glucuronate; triptorelin;tubulozole hydrochloride; uracil mustard; uredepa; vapreotide;verteporfin; vinblastine sulfate; vincristine sulfate; vindesine;vindesine sulfate; vinepidine sulfate; vinglycinate sulfate;vinleurosine sulfate; vinorelbine tartrate; vinrosidine sulfate;vinzolidine sulfate; vorozole; zeniplatin; zinostatin; and zorubicinhydrochloride.

Other anti-cancer drugs to be included within the methods orcompositions include, but are not limited to: 20-epi-1,25dihydroxyvitamin D3; 5-ethynyluracil; abiraterone; aclarubicin;acylfulvene; adecypenol; adozelesin; aldesleukin; ALL-TK antagonists;altretamine; ambamustine; amidox; amifostine; aminolevulinic acid;amrubicin; amsacrine; anagrelide; anastrozole; andrographolide;angiogenesis inhibitors; antagonist D; antagonist G; antarelix;anti-dorsalizing morphogenetic protein-1; antiandrogen, prostaticcarcinoma; antiestrogen; antineoplaston; antisense oligonucleotides;aphidicolin glycinate; apoptosis gene modulators; apoptosis regulators;apurinic acid; ara-CDP-DL-PTBA; arginine deaminase; asulacrine;atamestane; atrimustine; axinastatin 1; axinastatin 2; axinastatin 3;azasetron; azatoxin; azatyrosine; baccatin III derivatives; balanol;batimastat; BCR/ABL antagonists; benzochlorins; benzoylstaurosporine;beta lactam derivatives; beta-alethine; betaclamycin B; betulinic acid;bFGF inhibitor; bicalutamide; bisantrene; bisaziridinylspermine;bisnafide; bistratene A; bizelesin; breflate; bropirimine; budotitane;buthionine sulfoximine; calcipotriol; calphostin C; camptothecinderivatives; capecitabine; carboxamide-amino-triazole;carboxyamidotriazole; CaRest M3; CARN 700; cartilage derived inhibitor;carzelesin; casein kinase inhibitors (ICOS); castanospermine; cecropinB; cetrorelix; chlorins; chloroquinoxaline sulfonamide; cicaprost;cis-porphyrin; cladribine; clomifene analogues; clotrimazole;collismycin A; collismycin B; combretastatin A4; combretastatinanalogue; conagenin; crambescidin 816; crisnatol; cryptophycin 8;cryptophycin A derivatives; curacin A; cyclopentanthraquinones;cycloplatam; cypemycin; Ara-C ocfosfate; cytolytic factor; cytostatin;dacliximab; decitabine; dehydrodidemnin B; deslorelin; dexamethasone;dexifosfamide; dexrazoxane; dexverapamil; diaziquone; didemnin B; didox;diethylnorspermine; dihydro-5-azacytidine; dihydrotaxol, 9-; dioxamycin;diphenyl spiromustine; docetaxel; docosanol; dolasetron; doxifluridine;doxorubicin; droloxifene; dronabinol; duocarmycin SA; ebselen;ecomustine; edelfosine; edrecolomab; eflornithine; elemene; emitefur;epirubicin; epristeride; estramustine analogue; estrogen agonists;estrogen antagonists; etanidazole; etoposide phosphate; exemestane;fadrozole; fazarabine; fenretinide; filgrastim; finasteride;flavopiridol; flezelastine; fluasterone; fludarabine; fluorodaunorunicinhydrochloride; forfenimex; formestane; fostriecin; fotemustine;gadolinium texaphyrin; gallium nitrate; galocitabine; ganirelix;gelatinase inhibitors; gemcitabine; glutathione inhibitors; hepsulfam;heregulin; hexamethylene bisacetamide; hypericin; ibandronic acid;idarubicin; idoxifene; idramantone; ilmofosine; ilomastat; imatinib(e.g., Gleevee); imiquimod; immunostimulant peptides; insulin-likegrowth factor-1 receptor inhibitor; interferon agonists; interferons;interleukins; iobenguane; iododoxorubicin; ipomeanol, 4-; iroplact;irsogladine; isobengazole; isohomohalicondrin B; itasetron;jasplakinolide; kahalalide F; lamellarin-N triacetate; lanreotide;leinamycin; lenograstim; lentinan sulfate; leptolstatin; letrozole;leukemia inhibiting factor; leukocyte alpha interferon;leuprolide+estrogen+progesterone; leuprorelin; levamisole; liarozole;linear polyamine analogue; lipophilic disaccharide peptide; lipophilicplatinum compounds; lissoclinamide 7; lobaplatin; lombricine;lometrexol; lonidamine; losoxantrone; loxoribine; lurtotecan; lutetiumtexaphyrin; lysofylline; lytic peptides; maitansine; mannostatin A;marimastat; masoprocol; maspin; matrilysin inhibitors; matrixmetalloproteinase inhibitors; menogaril; merbarone; meterelin;methioninase; metoclopramide; MIF inhibitor; mifepristone; miltefosine;mirimostim; mitoguazone; mitolactol; mitomycin analogues; mitonafide;mitotoxin fibroblast growth factor-saporin; mitoxantrone; mofarotene;molgramostim; Erbitux, human chorionic gonadotrophin; monophosphoryllipid A+myobacterium cell wall sk; mopidamol; mustard anticancer agent;mycaperoxide B; mycobacterial cell wall extract; myriaporone;N-acetyldinaline; N-substituted benzamides; nafarelin; nagrestip;naloxone+pentazocine; napavin; naphterpin; nartograstim; nedaplatin;nemorubicin; neridronic acid; nilutamide; nisamycin; nitric oxidemodulators; nitroxide antioxidant; nitrullyn; oblimersen (Genasense®);O⁶-benzylguanine; octreotide; okicenone; oligonucleotides; onapristone;ondansetron; ondansetron; oracin; oral cytokine inducer; ormaplatin;osaterone; oxaliplatin; oxaunomycin; paclitaxel; paclitaxel analogues;paclitaxel derivatives; palauamine; palmitoylrhizoxin; pamidronic acid;panaxytriol; panomifene; parabactin; pazelliptine; pegaspargase;peldesine; pentosan polysulfate sodium; pentostatin; pentrozole;perflubron; perfosfamide; perillyl alcohol; phenazinomycin;phenylacetate; phosphatase inhibitors; picibanil; pilocarpinehydrochloride; pirarubicin; piritrexim; placetin A; placetin B;plasminogen activator inhibitor; platinum complex; platinum compounds;platinum-triamine complex; porfimer sodium; porfiromycin; prednisone;propyl bis-acridone; prostaglandin J2; proteasome inhibitors; proteinA-based immune modulator; protein kinase C inhibitor; protein kinase Cinhibitors, microalgal; protein tyrosine phosphatase inhibitors; purinenucleoside phosphorylase inhibitors; purpurins; pyrazoloacridine;pyridoxylated hemoglobin polyoxyethylene conjugate; raf antagonists;raltitrexed; ramosetron; ras farnesyl protein transferase inhibitors;ras inhibitors; ras-GAP inhibitor; retelliptine demethylated; rhenium Re186 etidronate; rhizoxin; ribozymes; RII retinamide; rohitukine;romurtide; roquinimex; rubiginone B1; ruboxyl; safingol; saintopin;SarCNU; sarcophytol A; sargramostim; Sdi 1 mimetics; semustine;senescence derived inhibitor 1; sense oligonucleotides; signaltransduction inhibitors; sizofiran; sobuzoxane; sodium borocaptate;sodium phenylacetate; solverol; somatomedin binding protein; sonermin;sparfosic acid; spicamycin D; spiromustine; splenopentin; spongistatin1; squalamine; stipiamide; stromelysin inhibitors; sulfinosine;superactive vasoactive intestinal peptide antagonist; suradista;suramin; swainsonine; tallimustine; tamoxifen methiodide; tauromustine;tazarotene; tecogalan sodium; tegafur; tellurapyrylium; telomeraseinhibitors; temoporfin; teniposide; tetrachlorodecaoxide; tetrazomine;thaliblastine; thiocoraline; thrombopoietin; thrombopoietin mimetic;thymalfasin; thymopoietin receptor agonist; thymotrinan; thyroidstimulating hormone; tin ethyl etiopurpurin; tirapazamine; titanocenebichloride; topsentin; toremifene; translation inhibitors; tretinoin;triacetyluridine; triciribine; trimetrexate; triptorelin; tropisetron;turosteride; tyrosine kinase inhibitors; tyrphostins; UBC inhibitors;ubenimex; urogenital sinus-derived growth inhibitory factor; urokinasereceptor antagonists; vapreotide; variolin B; velaresol; veramine;verdins; verteporfin; vinorelbine; vinxaltine; vitaxin; vorozole;zanoterone; zeniplatin; zilascorb; and zinostatin stimalamer.

In certain embodiments, a solid form of Compound 1 is administered incombination with checkpoint inhibitors. In one embodiment, onecheckpoint inhibitor is used in combination with a solid form ofCompound 1 in connection with the methods provided herein. In anotherembodiment, two checkpoint inhibitors are used in combination with asolid form of Compound 1 in connection with the methods provided herein.In yet another embodiment, three or more checkpoint inhibitors are usedin combination with a solid form of Compound 1 in connection with themethods provided herein.

As used herein, the term “immune checkpoint inhibitor” or “checkpointinhibitor” refers to molecules that totally or partially reduce,inhibit, interfere with or modulate one or more checkpoint proteins.Without being limited by a particular theory, checkpoint proteinsregulate T-cell activation or function. Numerous checkpoint proteins areknown, such as CTLA-4 and its ligands CD80 and CD86; and PD-1 with itsligands PD-L1 and PD-L2 (Pardoll, Nature Reviews Cancer, 2012, 12,252-264). These proteins appear responsible for co-stimulatory orinhibitory interactions of T-cell responses. Immune checkpoint proteinsappear to regulate and maintain self-tolerance and the duration andamplitude of physiological immune responses. Immune checkpointinhibitors include antibodies or are derived from antibodies.

In one embodiment, the checkpoint inhibitor is a CTLA-4 inhibitor. Inone embodiment, the CTLA-4 inhibitor is an anti-CTLA-4 antibody.Examples of anti-CTLA-4 antibodies include, but are not limited to,those described in U.S. Pat. Nos. 5,811,097; 5,811,097; 5,855,887;6,051,227; 6,207,157; 6,682,736; 6,984,720; and 7,605,238, all of whichare incorporated herein in their entireties. In one embodiment, theanti-CTLA-4 antibody is tremelimumab (also known as ticilimumab orCP-675,206). In another embodiment, the anti-CTLA-4 antibody isipilimumab (also known as MDX-010 or MDX-101). Ipilimumab is a fullyhuman monoclonal IgG antibody that binds to CTLA-4. Ipilimumab ismarketed under the trade name Yervoy™.

In one embodiment, the checkpoint inhibitor is a PD-1/PD-L1 inhibitor.Examples of PD-1/PD-L1 inhibitors include, but are not limited to, thosedescribed in U.S. Pat. Nos. 7,488,802; 7,943,743; 8,008,449; 8,168,757;8,217,149, and PCT Patent Application Publication Nos. WO2003042402,WO2008156712, WO2010089411, WO2010036959, WO2011066342, WO2011159877,WO2011082400, and WO2011161699, all of which are incorporated herein intheir entireties.

In one embodiment, the checkpoint inhibitor is a PD-1 inhibitor. In oneembodiment, the PD-1 inhibitor is an anti-PD-1 antibody. In oneembodiment, the anti-PD-1 antibody is nivolumab (also known as ONO-4538,BMS-936558, or MDX1106) or pembrolizumab (also known as MK-3475, SCH900475, or lambrolizumab). In one embodiment, the anti-PD-1 antibody isnivolumab. Nivolumab is a human IgG4 anti-PD-1 monoclonal antibody, andis marketed under the trade name Opdivo™. In another embodiment, theanti-PD-1 antibody is pembrolizumab. Pembrolizumab is a humanizedmonoclonal IgG4 antibody and is marketed under the trade name Keytruda™.In yet another embodiment, the anti-PD-1 antibody is CT-011, a humanizedantibody. CT-011 administered alone has failed to show response intreating acute myeloid leukemia (AML) at relapse. In yet anotherembodiment, the anti-PD-1 antibody is AMP-224, a fusion protein.

In one embodiment, the checkpoint inhibitor is a PD-L1 inhibitor. In oneembodiment, the PD-L1 inhibitor is an anti-PD-L1 antibody. In oneembodiment, the anti-PD-L1 antibody is MEDI4736 (durvalumab). In anotherembodiment, the anti-PD-L1 antibody is BMS-936559 (also known asMDX-1105-01). In yet another embodiment, the PD-L1 inhibitor isatezolizumab (also known as MPDL3280A, and Tecentriq®).

In one embodiment, the checkpoint inhibitor is a PD-L2 inhibitor. In oneembodiment, the PD-L2 inhibitor is an anti-PD-L2 antibody. In oneembodiment, the anti-PD-L2 antibody is rHIgM12B7A.

In one embodiment, the checkpoint inhibitor is a lymphocyte activationgene-3 (LAG-3) inhibitor. In one embodiment, the LAG-3 inhibitor isIMP321, a soluble Ig fusion protein (Brignone et al., J. Immunol., 2007,179, 4202-4211). In another embodiment, the LAG-3 inhibitor isBMS-986016.

In one embodiment, the checkpoint inhibitors is a B7 inhibitor. In oneembodiment, the B7 inhibitor is a B7-H3 inhibitor or a B7-H4 inhibitor.In one embodiment, the B7-H3 inhibitor is MGA271, an anti-B7-H3 antibody(Loo et al., Clin. Cancer Res., 2012, 3834).

In one embodiment, the checkpoint inhibitors is a TIM3 (T-cellimmunoglobulin domain and mucin domain 3) inhibitor (Fourcade et al., J.Exp. Med., 2010, 207, 2175-86; Sakuishi et al., J. Exp. Med., 2010, 207,2187-94).

In one embodiment, the checkpoint inhibitor is an OX40 (CD134) agonist.In one embodiment, the checkpoint inhibitor is an anti-OX40 antibody. Inone embodiment, the anti-OX40 antibody is anti-OX-40. In anotherembodiment, the anti-OX40 antibody is MEDI6469.

In one embodiment, the checkpoint inhibitor is a GITR agonist. In oneembodiment, the checkpoint inhibitor is an anti-GITR antibody. In oneembodiment, the anti-GITR antibody is TRX518.

In one embodiment, the checkpoint inhibitor is a CD137 agonist. In oneembodiment, the checkpoint inhibitor is an anti-CD137 antibody. In oneembodiment, the anti-CD137 antibody is urelumab. In another embodiment,the anti-CD137 antibody is PF-05082566.

In one embodiment, the checkpoint inhibitor is a CD40 agonist. In oneembodiment, the checkpoint inhibitor is an anti-CD40 antibody. In oneembodiment, the anti-CD40 antibody is CF-870,893.

In one embodiment, the checkpoint inhibitor is recombinant humaninterleukin-15 (rhIL-15).

In one embodiment, the checkpoint inhibitor is an IDO inhibitor. In oneembodiment, the IDO inhibitor is INCB024360. In another embodiment, theIDO inhibitor is indoximod.

In certain embodiments, the combination therapies provided hereininclude two or more of the checkpoint inhibitors described herein(including checkpoint inhibitors of the same or different class).Moreover, the combination therapies described herein can be used incombination with second active agents as described herein whereappropriate for treating diseases described herein and understood in theart.

In certain embodiments, Compound 1 can be used in combination with oneor more immune cells expressing one or more chimeric antigen receptors(CARs) on their surface (e.g., a modified immune cell). Generally, CARscomprise an extracellular domain from a first protein e.g., anantigen-binding protein), a transmembrane domain, and an intracellularsignaling domain. In certain embodiments, once the extracellular domainbinds to a target protein such as a tumor-associated antigen (TAA) ortumor-specific antigen (TSA), a signal is generated via theintracellular signaling domain that activates the immune cell, e.g., totarget and kill a cell expressing the target protein.

Extracellular domains: The extracellular domains of the CARs bind to anantigen of interest. In certain embodiments, the extracellular domain ofthe CAR comprises a receptor, or a portion of a receptor, that binds tosaid antigen. In certain embodiments, the extracellular domaincomprises, or is, an antibody or an antigen-binding portion thereof. Inspecific embodiments, the extracellular domain comprises, or is, asingle chain Fv (scFv) domain. The single-chain Fv domain can comprise,for example, a V_(L) linked to V_(H) by a flexible linker, wherein saidV_(L) and V_(H) are from an antibody that binds said antigen.

In certain embodiments, the antigen recognized by the extracellulardomain of a polypeptide described herein is a tumor-associated antigen(TAA) or a tumor-specific antigen (TSA). In various specificembodiments, the tumor-associated antigen or tumor-specific antigen is,without limitation, Her2, prostate stem cell antigen (PSCA),alpha-fetoprotein (AFP), carcinoembryonic antigen (CEA), cancerantigen-125 (CA-125), CA19-9, calretinin, MUC-1, B cell maturationantigen (BCMA), epithelial membrane protein (EMA), epithelial tumorantigen (ETA), tyrosinase, melanoma-24 associated antigen (MAGE), CD19,CD22, CD27, CD30, CD34, CD45, CD70, CD99, CD117, EGFRvIII (epidermalgrowth factor variant III), mesothelin, PAP (prostatic acidphosphatase), prostein, TARP (T cell receptor gamma alternate readingframe protein), Trp-p8, STEAPI (six-transmembrane epithelial antigen ofthe prostate 1), chromogranin, cytokeratin, desmin, glial fibrillaryacidic protein (GFAP), gross cystic disease fluid protein (GCDFP-15),HMB-45 antigen, protein melan-A (melanoma antigen recognized by Tlymphocytes; MART-I), myo-D1, muscle-specific actin (MSA),neurofilament, neuron-specific enolase (NSE), placental alkalinephosphatase, synaptophysis, thyroglobulin, thyroid transcriptionfactor-1, the dimeric form of the pyruvate kinase isoenzyme type M2(tumor M2-PK), an abnormal ras protein, or an abnormal p53 protein. Incertain other embodiments, the TAA or TSA recognized by theextracellular domain of a CAR is integrin αvβ3 (CD61), galactin, orRal-B.

In certain embodiments, the TAA or TSA recognized by the extracellulardomain of a CAR is a cancer/testis (CT) antigen, e.g., BAGE, CAGE,CTAGE, FATE, GAGE, HCA661, HOM-TES-85, MAGEA, MAGEB, MAGEC, NA88,NY-ESO-1, NY-SAR-35, OY-TES-1, SPANXBI, SPA17, SSX, SYCPI, or TPTE.

In certain other embodiments, the TAA or TSA recognized by theextracellular domain of a CAR is a carbohydrate or ganglioside, e.g.,fuc-GMI, GM2 (oncofetal antigen-immunogenic-1; OFA-I-1); GD2 (OFA-I-2),GM3, GD3, and the like.

In certain other embodiments, the TAA or TSA recognized by theextracellular domain of a CAR is alpha-actinin-4, Bage-1, BCR-ABL,Bcr-Abl fusion protein, beta-catenin, CA 125, CA 15-3 (CA 27.29\BCAA),CA 195, CA 242, CA-50, CAM43, Casp-8, cdc27, cdk4, cdkn2a, CEA, coa-1,dek-can fusion protein, EBNA, EF2, Epstein Barr virus antigens,ETV6-AML1 fusion protein, HLA-A2, HLA-All, hsp70-2, KIAA0205, Mart2,Mum-1, 2, and 3, neo-PAP, myosin class I, OS-9, pml-RARα fusion protein,PTPRK, K-ras, N-ras, triosephosphate isomerase, Gage 3,4,5,6,7, GnTV,Herv-K-mel, Lage-1, NA-88, NY-Eso-1/Lage-2, SP17, SSX-2, TRP2-Int2,gp100 (Pme117), tyrosinase, TRP-1, TRP-2, MAGE-1, MAGE-3, RAGE, GAGE-1,GAGE-2, p15(58), RAGE, SCP-1, Hom/Me1-40, PRAME, p53, HRas, HER-2/neu,E2A-PRL, H4-RET, IGH-IGK, MYL-RAR, human papillomavirus (HPV) antigensE6 and E7, TSP-180, MAGE-4, MAGE-5, MAGE-6, p185erbB2, p180erbB-3,c-met, nm-23H1, PSA, TAG-72-4, CA 19-9, CA 72-4, CAM 17.1, NuMa, K-ras,13-Catenin, Mum-1, p16, TAGE, PSMA, CT7, telomerase, 43-9F, 5T4,791Tgp72, 13HCG, BCA225, BTAA, CD68\KP1, CO-029, FGF-5, G250, Ga733(EpCAM), HTgp-175, M344, MA-50, MG7-Ag, MOV18, NB\70K, NY-C0-1, RCAS1,SDCCAG16, TA-90, TAAL6, TAG72, TLP, or TPS.

In various specific embodiments, the tumor-associated antigen ortumor-specific antigen is an AML-related tumor antigens, as described inS. Anguille et al, Leukemia (2012), 26, 2186-2196.

Other tumor-associated and tumor-specific antigens are known to those inthe art.

Receptors, antibodies, and scFvs that bind to TSAs and TAAs, useful inconstructing chimeric antigen receptors, are known in the art, as arenucleotide sequences that encode them.

In certain specific embodiments, the antigen recognized by theextracellular domain of a chimeric antigen receptor is an antigen notgenerally considered to be a TSA or a TAA, but which is neverthelessassociated with tumor cells, or damage caused by a tumor. In certainembodiments, for example, the antigen is, e.g., a growth factor,cytokine or interleukin, e.g., a growth factor, cytokine, or interleukinassociated with angiogenesis or vasculogenesis. Such growth factors,cytokines, or interleukins can include, e.g., vascular endothelialgrowth factor (VEGF), basic fibroblast growth factor (bFGF),platelet-derived growth factor (PDGF), hepatocyte growth factor (HGF),insulin-like growth factor (IGF), or interleukin-8 (IL-8). Tumors canalso create a hypoxic environment local to the tumor. As such, in otherspecific embodiments, the antigen is a hypoxia-associated factor, e.g.,HIF-1α, HIF-2α, HIF-2β, HIF-3α, or HIF-3β. Tumors can also causelocalized damage to normal tissue, causing the release of moleculesknown as damage associated molecular pattern molecules (DAMPs; alsoknown as alarmins). In certain other specific embodiments, therefore,the antigen is a DAMP, e.g., a heat shock protein, chromatin-associatedprotein high mobility group box 1 (HMGB 1), S100A8 (MRP8, calgranulinA), S100A9 (MRP14, calgranulin B), serum amyloid A (SAA), or can be adeoxyribonucleic acid, adenosine triphosphate, uric acid, or heparinsulfate.

Transmembrane domain: In certain embodiments, the extracellular domainof the CAR is joined to the transmembrane domain of the polypeptide by alinker, spacer or hinge polypeptide sequence, e.g., a sequence from CD28or a sequence from CTLA4. The transmembrane domain can be obtained orderived from the transmembrane domain of any transmembrane protein, andcan include all or a portion of such transmembrane domain. In specificembodiments, the transmembrane domain can be obtained or derived from,e.g., CD8, CD16, a cytokine receptor, and interleukin receptor, or agrowth factor receptor, or the like.

Intracellular signaling domains: In certain embodiments, theintracellular domain of a CAR is or comprises an intracellular domain ormotif of a protein that is expressed on the surface of T cells andtriggers activation and/or proliferation of said T cells. Such a domainor motif is able to transmit a primary antigen-binding signal that isnecessary for the activation of a T lymphocyte in response to theantigen's binding to the CAR's extracellular portion. Typically, thisdomain or motif comprises, or is, an ITAM (immunoreceptor tyrosine-basedactivation motif). ITAM-containing polypeptides suitable for CARsinclude, for example, the zeta CD3 chain (CD3) or ITAM-containingportions thereof. In a specific embodiment, the intracellular domain isa CD3ζ intracellular signaling domain. In other specific embodiments,the intracellular domain is from a lymphocyte receptor chain, a TCR/CD3complex protein, an Fe receptor subunit or an IL-2 receptor subunit. Incertain embodiments, the CAR additionally comprises one or moreco-stimulatory domains or motifs, e.g., as part of the intracellulardomain of the polypeptide. The one or more co-stimulatory domains ormotifs can be, or can comprise comprise, one or more of a co-stimulatoryCD27 polypeptide sequence, a co-stimulatory CD28 polypeptide sequence, aco-stimulatory OX40 (CD134) polypeptide sequence, a co-stimulatory 4-1BB(CD137) polypeptide sequence, or a co-stimulatory inducible T-cellcostimulatory (ICOS) polypeptide sequence, or other costimulatory domainor motif, or any combination thereof.

The CAR may also comprise a T cell survival motif. The T cell survivalmotif can be any polypeptide sequence or motif that facilitates thesurvival of the T lymphocyte after stimulation by an antigen. In certainembodiments, the T cell survival motif is, or is derived from, CD3,CD28, an intracellular signaling domain of IL-7 receptor (IL-7R), anintracellular signaling domain of IL-12 receptor, an intracellularsignaling domain of IL-15 receptor, an intracellular signaling domain ofIL-21 receptor, or an intracellular signaling domain of transforminggrowth factor 0 (TGFβ) receptor.

The modified immune cells expressing the CARs can be, e.g., Tlymphocytes (T cells, e.g., CD4+ T cells or CD8+ T cells), cytotoxiclymphocytes (CTLs) or natural killer (NK) cells. T lymphocytes used inthe compositions and methods provided herein may be naive T lymphocytesor MHC-restricted T lymphocytes. In certain embodiments, the Tlymphocytes are tumor infiltrating lymphocytes (TILs). In certainembodiments, the T lymphocytes have been isolated from a tumor biopsy,or have been expanded from T lymphocytes isolated from a tumor biopsy.In certain other embodiments, the T cells have been isolated from, orare expanded from T lymphocytes isolated from, peripheral blood, cordblood, or lymph. Immune cells to be used to generate modified immunecells expressing a CAR can be isolated using art-accepted, routinemethods, e.g., blood collection followed by apheresis and optionallyantibody-mediated cell isolation or sorting.

The modified immune cells are preferably autologous to an individual towhom the modified immune cells are to be administered. In certain otherembodiments, the modified immune cells are allogeneic to an individualto whom the modified immune cells are to be administered. Whereallogeneic T lymphocytes or NK cells are used to prepare modified Tlymphocytes, it is preferable to select T lymphocytes or NK cells thatwill reduce the possibility of graft-versus-host disease (GVHD) in theindividual. For example, in certain embodiments, virus-specific Tlymphocytes are selected for preparation of modified T lymphocytes; suchlymphocytes will be expected to have a greatly reduced native capacityto bind to, and thus become activated by, any recipient antigens. Incertain embodiments, recipient-mediated rejection of allogeneic Tlymphocytes can be reduced by co-administration to the host of one ormore immunosuppressive agents, e.g., cyclosporine, tacrolimus,sirolimus, cyclophosphamide, or the like.

T lymphocytes, e.g., unmodified T lymphocytes, or T lymphocytesexpressing CD3 and CD28, or comprising a polypeptide comprising a CD3tsignaling domain and a CD28 co-stimulatory domain, can be expanded usingantibodies to CD3 and CD28, e.g., antibodies attached to beads; see,e.g., U.S. Pat. Nos. 5,948,893; 6,534,055; 6,352,694; 6,692,964;6,887,466; and 6,905,681.

The modified immune cells, e.g., modified T lymphocytes, can optionallycomprise a “suicide gene” or “safety switch” that enables killing ofsubstantially all of the modified immune cells when desired. Forexample, the modified T lymphocytes, in certain embodiments, cancomprise an HSV thymidine kinase gene (HSV-TK), which causes death ofthe modified T lymphocytes upon contact with gancyclovir. In anotherembodiment, the modified T lymphocytes comprise an inducible caspase,e.g., an inducible caspase 9 (icaspase9), e.g., a fusion protein betweencaspase 9 and human FK506 binding protein allowing for dimerizationusing a specific small molecule pharmaceutical. See Straathof et al.,Blood 1 05(11):4247-4254 (2005).

Specific second active agents particularly useful in the methods orcompositions include, but are not limited to, rituximab, oblimersen(Genasense®), remicade, docetaxel, celecoxib, melphalan, dexamethasone(Decadron®), steroids, gemcitabine, cisplatinum, temozolomide,etoposide, cyclophosphamide, temodar, carboplatin, procarbazine,gliadel, tamoxifen, topotecan, methotrexate, Arisa®, taxol, taxotere,fluorouracil, leucovorin, irinotecan, xeloda, interferon alpha,pegylated interferon alpha (e.g., PEG INTRON-A), capecitabine,cisplatin, thiotepa, fludarabine, carboplatin, liposomal daunorubicin,Ara-C, doxetaxol, pacilitaxel, vinblastine, IL-2, GM-CSF, dacarbazine,vinorelbine, zoledronic acid, palmitronate, biaxin, busulphan,prednisone, bisphosphonate, arsenic trioxide, vincristine, doxorubicin(Doxil®), paclitaxel, ganciclovir, adriamycin, estramustine sodiumphosphate (Emcyt®), sulindac, and etoposide.

In certain embodiments of the methods provided herein, use of a secondactive agent in combination with a solid form of Compound 1 providedherein may be modified or delayed during or shortly followingadministration of a solid form of Compound 1 provided herein as deemedappropriate by the practitioner of skill in the art. In certainembodiments, subjects being administered a solid form of Compound 1provided herein alone or in combination with other therapies may receivesupportive care including antiemetics, myeloid growth factors, andtransfusions of platelets, when appropriate. In some embodiments,subjects being administered a solid form of Compound 1 provided hereinmay be administered a growth factor as a second active agent accordingto the judgment of the practitioner of skill in the art. In someembodiments, provided is administration of a solid form of Compound 1provided herein in combination with erythropoietin or darbepoetin(Aranesp).

In certain embodiments, a solid form of Compound 1 provided herein isadministered with gemcitabine, cisplatinum, 5-fluorouracil, mitomycin,methotrexate, vinblastine, doxorubicin, carboplatin, thiotepa,paclitaxel or docetaxel to patients with locally advanced or metastatictransitional cell bladder cancer.

In certain embodiments, a solid form of Compound 1 provided herein isadministered in combination with a second active ingredient as follows:temozolomide to pediatric patients with relapsed or progressive braintumors or recurrent neuroblastoma; celecoxib, etoposide andcyclophosphamide for relapsed or progressive CNS cancer; temodar topatients with recurrent or progressive meningioma, malignant meningioma,hemangiopericytoma, multiple brain metastases, relapased brain tumors,or newly diagnosed glioblastoma multiforms; irinotecan to patients withrecurrent glioblastoma; carboplatin to pediatric patients with brainstem glioma; procarbazine to pediatric patients with progressivemalignant gliomas; cyclophosphamide to patients with poor prognosismalignant brain tumors, newly diagnosed or recurrent glioblastomamultiforms; Gliadel® for high grade recurrent malignant gliomas;temozolomide and tamoxifen for anaplastic astrocytoma; or topotecan forgliomas, glioblastoma, anaplastic astrocytoma or anaplasticoligodendroglioma.

In certain embodiments, a solid form of Compound 1 provided herein isadministered with methotrexate, cyclophosphamide, 5-fluorouracil,taxane, everolimus, abraxane, lapatinib, herceptin, pamidronatedisodium, eribulin mesylate, everolimus, gemcitabine, palbociclib,ixabepilone, kadcyla, pertuzumab, theotepa, aromatase inhibitors,exemestane, selective estrogen modulators, estrogen receptorantagonists, anthracyclines, emtansine, and/or pexidartinib to patientswith metastatic breast cancer.

In certain embodiments, a solid form of Compound 1 provided herein isadministered with temozolomide, doxorubicin (Adriamycin), fluorouracil(Adrucil, 5-fluorouracil), or streptozocin (Zanosar) to patients withneuroendocrine tumors.

In certain embodiments, a solid form of Compound 1 provided herein isadministered with methotrexate, gemcitabine, cisplatin, cetuximab,5-fluorouracil, bleomycin, docetaxel or carboplatin to patients withrecurrent or metastatic head or neck cancer.

In certain embodiments, a solid form of Compound 1 provided herein isadministered with gemcitabine, abraxane, 5-fluorouracil, afinitor,irinotecan, mitomycin C, sunitinib or tarceva to patients withpancreatic cancer.

In certain embodiments, a solid form of Compound 1 provided herein isadministered to patients with colon cancer in combination with ARISA®,avastatin, oxaliplatin, 5-fluorouracil, irinotecan, capecitabine,cetuximab, ramucirumab, panitumumab, bevacizumab, leucovorin calcium,lonsurf, regorafenib, ziv-aflibercept, taxol, and/or taxotere.

In certain embodiments, a solid form of Compound 1 provided herein isadministered with capecitabine and/or vemurafenib to patients withrefractory colorectal cancer or patients who fail first line therapy orhave poor performance in colon or rectal adenocarcinoma.

In certain embodiments, a solid form of Compound 1 provided herein isadministered in combination with fluorouracil, leucovorin, andirinotecan to patients with Dukes C & D colorectal cancer or to patientswho have been previously treated for metastatic colorectal cancer.

In certain embodiments, a solid form of Compound 1 provided herein isadministered to patients with refractory colorectal cancer incombination with capecitabine, xeloda, and/or irinotecan.

In certain embodiments, a solid form of Compound 1 provided herein isadministered with capecitabine and irinotecan to patients withrefractory colorectal cancer or to patients with unresectable ormetastatic colorectal carcinoma.

In certain embodiments, a solid form of Compound 1 provided herein isadministered alone or in combination with interferon alpha orcapecitabine to patients with unresectable or metastatic hepatocellularcarcinoma; or with cisplatin and thiotepa, or with sorafenib tosylate topatients with primary or metastatic liver cancer.

In certain embodiments, a solid form of Compound 1 provided herein isadministered in combination with doxorubicin, paclitaxel, vinblastine orpegylated interferon alpha to patients with Kaposi's sarcoma.

In certain embodiments, a solid form of Compound 1 provided herein isadministered in combination with arsenic trioxide, fludarabine,carboplatin, daunorubicin, cyclophosphamide, cytarabine, doxorubicin,idarubicin, mitoxantrone hydrochloride, thioguanine, vincritine, and/ortopotecan to patients with refractory or relapsed or high-risk acutemyeloid leukemia.

In certain embodiments, a solid form of Compound 1 provided herein isadministered in combination with liposomal daunorubicin, topotecanand/or cytarabine to patients with unfavorable karotype acutemyeloblastic leukemia.

In certain embodiments, a solid form of Compound 1 provided herein isadministered in combination with methotrexate, mechlorethaminehydrochloride, afatinib dimaleate, pemetrexed, bevacizumab, carboplatin,cisplatin, ceritinib, crizotinib, ramucirumab, pembrolizumab, docetaxel,vinorelbine tartrate, gemcitabine, abraxane, erlotinib, geftinib, and/oririnotecan to patients with non-small cell lung cancer.

In certain embodiments, a solid form of Compound 1 provided herein isadministered in combination with carboplatin and irinotecan to patientswith non-small cell lung cancer.

In certain embodiments, a solid form of Compound 1 provided herein isadministered with doxetaxol to patients with non-small cell lung cancerwho have been previously treated with carbo/etoposide and radiotherapy.

In certain embodiments, a solid form of Compound 1 provided herein isadministered in combination with carboplatin and/or taxotere, or incombination with carboplatin, pacilitaxel and/or thoracic radiotherapyto patients with non-small cell lung cancer.

In certain embodiments, a solid form of Compound 1 provided herein isadministered in combination with taxotere to patients with stage IIIB orIV non-small cell lung cancer.

In certain embodiments, a solid form of Compound 1 provided herein isadministered in combination with oblimersen (Genasense®), methotrexate,mechlorethamine hydrochloride, etoposide, topotecan or doxorubicin topatients with small cell lung cancer.

In certain embodiments, a solid form of Compound 1 provided herein isadministered in combination with ABT-737 (Abbott Laboratories) and/orobatoclax (GX15-070) to patients with lymphoma and other blood cancers.

In certain embodiments, a solid form of Compound 1 provided herein isadministered alone or in combination with a second active ingredientsuch as vinblastine or fludarabine adcetris, ambochlorin, becenum,bleomycin, brentuximab vedotin, carmustinem chlorambucil,cyclophosphamide, dacarbazine, doxorubicin, lomustine, matulane,mechlorethamine hydrochloride, prednisone, procarbazine hydrochloride orvincristine to patients with various types of lymphoma, including, butnot limited to, Hodgkin's lymphoma, non-Hodgkin's lymphoma, cutaneousT-Cell lymphoma, cutaneous B-Cell lymphoma, diffuse large B-Celllymphoma or relapsed or refractory low grade follicular lymphoma.

In certain embodiments, a solid form of Compound 1 provided herein isadministered in combination with taxotere, dabrafenib, imlygic,ipilimumab, pembrolizumab, nivolumab, trametinib, vemurafenib,talimogene laherparepvec, IL-2, IFN, GM-CSF, and/or dacarbazine topatients with various types or stages of melanoma.

In certain embodiments, a solid form of Compound 1 provided herein isadministered alone or in combination with vinorelbine to patients withmalignant mesothelioma, or stage IIIB non-small cell lung cancer withpleural implants or malignant pleural effusion mesothelioma syndrome.

In certain embodiments, a solid form of Compound 1 provided herein isadministered to patients with various types or stages of multiplemyeloma in combination with dexamethasone, zoledronic acid,palmitronate, GM-CSF, biaxin, vinblastine, melphalan, busulphan,cyclophosphamide, IFN, prednisone, bisphosphonate, celecoxib, arsenictrioxide, PEG INTRON-A, vincristine, becenum, bortezomib, carfilzomib,doxorubicin, panobinostat, lenalidomide, pomalidomide, thalidomide,mozobil or a combination thereof.

In certain embodiments, a solid form of Compound 1 provided herein isadministered to patients with various types or stages of multiplemyeloma in combination with chimeric antigen receptor (CAR) T-cells.

In certain embodiments, a solid form of Compound 1 provided herein isadministered to patients with relapsed or refractory multiple myeloma incombination with doxorubicin (Doxir®), vincristine and/or dexamethasone(Decadron®).

In certain embodiments, a solid form of Compound 1 provided herein isadministered to patients with various types or stages of ovarian cancersuch as peritoneal carcinoma, papillary serous carcinoma, refractoryovarian cancer or recurrent ovarian cancer, in combination with taxol,carboplatin, doxorubicin, gemcitabine, cisplatin, xeloda, paclitaxel,dexamethasone, avastin, cyclophosphamide, topotecan, olaparib, thiotepa,or a combination thereof.

In certain embodiments, a solid form of Compound 1 provided herein isadministered to patients with various types or stages of prostatecancer, in combination with xeloda, 5 FU/LV, gemcitabine, irinotecanplus gemcitabine, cyclophosphamide, vincristine, dexamethasone, GM-CSF,celecoxib, taxotere, ganciclovir, paclitaxel, adriamycin, docetaxel,estramustine, Emcyt, denderon, zytiga, bicalutamide, cabazitaxel,degarelix, enzalutamide, zoladex, leuprolide acetate, mitoxantronehydrochloride, prednisone, sipuleucel-T, radium 223 dichloride, or acombination thereof.

In certain embodiments, a solid form of Compound 1 provided herein isadministered to patients with various types or stages of renal cellcancer, in combination with capecitabine, IFN, tamoxifen, IL-2, GM-CSF,Celebrex®, or a combination thereof.

In certain embodiments, a solid form of Compound 1 provided herein isadministered to patients with various types or stages of gynecologic,uterus or soft tissue sarcoma cancer in combination with IFN,dactinomycin, doxorubicin, imatinib mesylate, pazopanib, hydrochloride,trabectedin, a COX-2 inhibitor such as Celebrex®, and/or sulindac.

In certain embodiments, a solid form of Compound 1 provided herein isadministered to patients with various types or stages of solid tumors incombination with celebrex, etoposide, cyclophosphamide, docetaxel,apecitabine, IFN, tamoxifen, IL-2, GM-CSF, or a combination thereof.

In certain embodiments, a solid form of Compound 1 provided herein isadministered to patients with scleroderma or cutaneous vasculitis incombination with celebrex, etoposide, cyclophosphamide, docetaxel,apecitabine, IFN, tamoxifen, IL-2, GM-CSF, or a combination thereof.

In certain embodiments, a solid form of Compound 1 provided herein isadministered to patients with MDS in combination with azacitidine,cytarabine, daunorubicin, decitabine, idarubicin, lenalidomide or acombination thereof.

Also encompassed herein is a method of increasing the dosage of ananti-cancer drug or agent that can be safely and effectivelyadministered to a patient, which comprises administering to the patient(e.g., a human) a solid form of Compound 1 provided herein. Patientsthat can benefit by this method are those likely to suffer from anadverse effect associated with anti-cancer drugs for treating a specificcancer of the skin, subcutaneous tissue, lymph nodes, brain, lung,liver, bone, intestine, colon, heart, pancreas, adrenal, kidney,prostate, breast, colorectal, or combinations thereof. Theadministration of a solid form of Compound 1 provided herein alleviatesor reduces adverse effects which are of such severity that it wouldotherwise limit the amount of anti-cancer drug.

In one embodiment, a solid form of Compound 1 provided herein isadministered daily in an amount ranging from about 0.1 to about 150 mg,from about 1 to about 50 mg, or from about 2 to about 25 mg, or fromabout 1 to about 10 mg, prior to, during, or after the occurrence of theadverse effect associated with the administration of an anti-cancer drugto a patient. In certain embodiments, a solid form of Compound 1provided herein is administered in combination with specific agents suchas heparin, aspirin, coumadin, or G-CSF to avoid adverse effects thatare associated with anti-cancer drugs such as but not limited toneutropenia or thrombocytopenia.

In one embodiment, a solid form of Compound 1 provided herein isadministered to patients with diseases and disorders associated with orcharacterized by, undesired angiogenesis in combination with additionalactive ingredients, including, but not limited to, anti-cancer drugs,anti-inflammatories, antihistamines, antibiotics, and steroids.

In another embodiment, encompassed herein is a method of treating,preventing and/or managing cancer, which comprises administering a solidform of Compound 1 provided herein in conjunction with (e.g. before,during, or after) conventional therapy including, but not limited to,surgery, immunotherapy, biological therapy, radiation therapy, or othernon-drug based therapy presently used to treat, prevent and/or managecancer. The combined use of the compound provided herein andconventional therapy may provide a unique treatment regimen that isunexpectedly effective in certain patients. Without being limited bytheory, it is believed that a solid form of Compound 1 provided hereinmay provide additive or synergistic effects when given concurrently withconventional therapy.

As discussed elsewhere herein, encompassed herein is a method ofreducing, treating and/or preventing adverse or undesired effectsassociated with conventional therapy including, but not limited to,surgery, chemotherapy, radiation therapy, hormonal therapy, biologicaltherapy and immunotherapy. A solid form of Compound 1 provided hereinand other active ingredient can be administered to a patient prior to,during, or after the occurrence of the adverse effect associated withconventional therapy.

In certain embodiments, the methods provided herein compriseadministration of calcium, calcitriol, and vitamin D supplementationwith a solid form of Compound 1. In certain embodiments, the methodsprovided herein comprise administration of calcium, calcitriol, andvitamin D supplementation prior to the treatment with a solid form ofCompound 1.

In certain embodiments, calcium supplementation is administered todeliver at least 1200 mg of elemental calcium per day given in divideddoses. In certain embodiments, calcium supplementation is administeredas calcium carbonate in a dose of 500 mg administered three times a dayper orally (PO).

In certain embodiments, calcitriol supplementation is administered todeliver 0.25 μg calcitriol (PO) once daily.

In certain embodiments, vitamin D supplementation is administered todeliver about 500 IU to about 5000 IU vitamin D once daily. In certainembodiments, vitamin D supplementation is administered to deliver about1000 IU vitamin D once daily. In certain embodiments, vitamin Dsupplementation is administered to deliver about 50,000 IU vitamin Dweekly. In certain embodiments, vitamin D supplementation isadministered to deliver about 1000 IU vitamin D2 or D3 once daily. Incertain embodiments, vitamin D supplementation is administered todeliver about 50,000 IU vitamin D2 or D3 weekly.

In one embodiment, the solid form of Compound 1 can be administered inan amount ranging from about 0.1 to about 150 mg, from about 1 to about25 mg, or from about 2 to about 10 mg daily alone, or in combinationwith a second active agent disclosed herein, prior to, during, or afterthe use of conventional therapy.

In certain embodiments, a solid form of Compound 1 provided herein anddoxetaxol are administered to patients with non-small cell lung cancerwho were previously treated with carbo/VP 16 and radiotherapy.

6.3.2. Use With Transplantation Therapy

The solid forms of Compound 1 provided herein provided herein can beused to reduce the risk of Graft Versus Host Disease (GVHD). Therefore,encompassed herein is a method of treating, preventing and/or managingcancer, which comprises administering a solid form of Compound 1provided herein in conjunction with transplantation therapy.

As those of ordinary skill in the art are aware, the treatment of canceris often based on the stages and mechanism of the disease. For example,as inevitable leukemic transformation develops in certain stages ofcancer, transplantation of peripheral blood stem cells, hematopoieticstem cell preparation or bone marrow may be necessary. The combined useof a solid form of Compound 1 provided herein provided herein andtransplantation therapy provides a unique and unexpected synergism. Inparticular, a solid form of Compound 1 provided herein exhibitsimmunomodulatory activity that may provide additive or synergisticeffects when given concurrently with transplantation therapy in patientswith cancer.

The solid forms of Compound 1 provided herein can work in combinationwith transplantation therapy reducing complications associated with theinvasive procedure of transplantation and risk of GVHD. Encompassedherein is a method of treating, preventing and/or managing cancer whichcomprises administering to a patient (e.g., a human) solid form ofCompound 1 provided herein before, during, or after the transplantationof umbilical cord blood, placental blood, peripheral blood stem cell,hematopoietic stem cell preparation, or bone marrow. Some examples ofstem cells suitable for use in the methods provided herein are disclosedin U.S. Pat. No. 7,498,171, the disclosure of which is incorporatedherein by reference in its entirety.

In one embodiment, a solid form of Compound 1 provided herein isadministered to patients with acute myeloid leukemia before, during, orafter transplantation.

In one embodiment, a solid form of Compound 1 provided herein isadministered to patients with multiple myeloma before, during, or afterthe transplantation of autologous peripheral blood progenitor cell.

In one embodiment, a solid form of Compound 1 provided herein isadministered to patients with NHL (e.g., DLBCL) before, during, or afterthe transplantation of autologous peripheral blood progenitor cell.

6.3.3. Cycling Therapy

In certain embodiments, the prophylactic or therapeutic agents providedherein are cyclically administered to a patient. Cycling therapyinvolves the administration of an active agent for a period of time,followed by a rest for a period of time, and repeating this sequentialadministration. Cycling therapy can reduce the development of resistanceto one or more of the therapies, avoid, or reduce the side effects ofone of the therapies, and/or improves the efficacy of the treatment.

Consequently, in certain embodiments, a solid form of Compound 1provided herein provided herein is administered daily in a single ordivided doses in a four to six week cycle with a rest period of about aweek or two weeks. The cycling method further allows the frequency,number, and length of dosing cycles to be increased. Thus, encompassedherein in certain embodiments is the administration of a solid form ofCompound 1 provided herein for more cycles than are typical when it isadministered alone. In certain embodiments, a solid form of Compound 1provided herein is administered for a greater number of cycles thatwould typically cause dose-limiting toxicity in a patient to whom asecond active ingredient is not also being administered.

In one embodiment, a solid form of Compound 1 provided herein isadministered daily and continuously for three or four weeks at a dose offrom about 0.1 to about 150 mg/d followed by a break of one or twoweeks. In another embodiment, a solid form of Compound 1 provided hereinis administered once a day for the first two to five or ten days of a 28day cycle, followed by a rest period with no administration for the restof the 28 day cycle in a dose from about 1 to about 10 mg.

In another embodiment, a solid form of Compound 1 provided herein and asecond active ingredient are administered orally, with administration ofthe solid form of Compound 1 occurring 30 to 60 minutes prior to asecond active ingredient, during a cycle of four to six weeks. Incertain embodiments, the combination of a solid form of Compound 1provided herein and a second active ingredient is administered byintravenous infusion over about 90 minutes every cycle. In certainembodiments, one cycle comprises the administration from about 0.1 toabout 150 mg/day of a solid form of Compound 1 provided herein and fromabout 50 to about 200 mg/m²/day of a second active ingredient daily forthree to four weeks and then one or two weeks of rest. In certainembodiments, the number of cycles during which the combinatorialtreatment is administered to a patient is ranging from about one toabout 24 cycles, from about two to about 16 cycles, or from about fourto about three cycles.

6.4. Patient Population

In certain embodiments of the methods provided herein, the subject is ananimal, preferably a mammal, more preferably a non-human primate. Inparticular embodiments, the subject is a human. The subject can be amale or female subject.

Particularly useful subjects for the methods provided herein includehuman cancer patients, for example, those who have been diagnosed withleukemia, including acute myeloid leukemia, acute lymphocytic leukemia,chronic myelogenous leukemia, and chronic myelogenous leukemia. Incertain embodiments, the subject has not been diagnosed with acutepromyelocytic leukemia.

In some embodiments, the subject has a higher than normal blastpopulation. In some embodiments, the subject has a blast population ofat least 10%. In some embodiments, the subject has a blast population ofbetween 10 and 15%. In some embodiments, the subject has a blastpopulation of at least 15%. In some embodiments, the subject has a blastpopulation of between 15 and 20%. In some embodiments, the subject has ablast population of at least 20%. In some embodiments, the subject has ablast population of about 10-15%, about 15-20%, or about 20-25%. Inother embodiments, the subject has a blast population of less than 10%.In the context of the methods described herein, useful subjects having ablast population of less than 10% includes those subjects that, for anyreason according to the judgment of the skilled practitioner in the art,are in need of treatment with a compound provided herein, alone or incombination with a second active agent.

In some embodiments, the subject is treated based on the EasternCooperative Oncology Group (ECOG) performance status score of thesubject for leukemia. ECOG performance status can be scored on a scaleof 0 to 5, with 0 denoting asymptomatic; 1 denoting symptomatic butcompletely ambulant; 2 denoting symptomatic and <50% in bed during theday; 3 denoting symptomatic and >50% in bed, but not bed bound; 4denoting bed bound; and 5 denoting death. In some embodiments, thesubject has an ECOG performance status score of 0 or 1. In someembodiments, the subject has an ECOG performance status score of 0. Insome embodiments, the subject has an ECOG performance status score of 1.In other embodiments, the subject has an ECOG performance status scoreof 2.

In certain embodiments, the methods provided herein encompass thetreatment of subjects who have not been previously treated for leukemia.In some embodiments, the subject has not undergone allogeneic bonemarrow transplantation. In some embodiments, the subject has notundergone a stem cell transplantation. In some embodiments, the subjecthas not received hydroxyurea treatment. In some embodiments, the subjecthas not been treated with any investigational products for leukemia. Insome embodiments, the subject has not been treated with systemicglucocorticoids.

In other embodiments, the methods encompass treating subjects who havebeen previously treated or are currently being treated for leukemia. Forexample, the subject may have been previously treated or are currentlybeing treated with a standard treatment regimen for leukemia. Thesubject may have been treated with any standard leukemia treatmentregimen known to the practitioner of skill in the art. In certainembodiments, the subject has been previously treated with at least oneinduction/reinduction or consolidation AML regimen. In some embodiments,the subject has undergone autologous bone marrow transplantation or stemcell transplantation as part of a consolidation regimen. In someembodiments, the bone marrow or stem cell transplantation occurred atleast 3 months prior to treatment according to the methods providedherein. In some embodiments, the subject has undergone hydroxyureatreatment. In some embodiments, the hydroxyurea treatment occurred nolater than 24 hours prior to treatment according to the methods providedherein. In some embodiments, the subject has undergone prior inductionor consolidation therapy with cytarabine (Ara-C). In some embodiments,the subject has undergone treatment with systemic glucocorticosteroids.In some embodiments, the glucocorticosteroid treatment occurred no later24 hours prior to treatment according to the methods described herein.In other embodiments, the methods encompass treating subjects who havebeen previously treated for cancer, but are non-responsive to standardtherapies.

Also encompassed are methods of treating subjects having relapsed orrefractory leukemia. In some embodiments, the subject has been diagnosedwith a relapsed or refractory AML subtype, as defined by the WorldHealth Organization (WHO). Relapsed or refractory disease may be de novoAML or secondary AML, e.g., therapy-related AML (t-AML).

In some embodiments, the methods provided herein are used to treat drugresistant leukemias, such as chronic myelogenous leukemia (CML). Thus,treatment with a compound provided herein could provide an alternativefor patients who do not respond to other methods of treatment. In someembodiments, such other methods of treatment encompass treatment withGleevec® (imatinib mesylate). In some embodiments, provided herein aremethods of treatment of Philadelphia chromosome positive chronicmyelogenous leukemia (Ph+CML). In some embodiments, provided herein aremethods of treatment of Gleevec® (imatinib mesylate) resistantPhiladelphia chromosome positive chronic myelogenous leukemia (Ph+CML).

Also encompassed are methods of treating a subject regardless of thesubject's age, although some diseases or disorders are more common incertain age groups. In some embodiments, the subject is at least 18years old. In some embodiments, the subject is more than 18, 25, 35, 40,45, 50, 55, 60, 65, or 70 years old. In other embodiments, the subjectis less than 65 years old. In some embodiments, the subject is less than18 years old. In some embodiments, the subject is less than 18, 15, 12,10, 9, 8 or 7 years old.

In some embodiments, the methods may find use in subjects at least 50years of age, although younger subjects could benefit from the method aswell. In other embodiments, the subjects are at least 55, at least 60,at least 65, and at least 70 years of age. In another embodiment, thesubjects have adverse cytogenetics. “Adverse cytogenetics” is defined asany nondiploid karyotype, or greater than or equal to 3 chromosomalabnormalities. In another embodiment, the subjects are at least 60 yearsof age and have adverse cytogenetics. In another embodiment, thesubjects are 60-65 years of age and have adverse cytogenetics. Inanother embodiment, the subjects are 65-70 years of age and have adversecytogenetics.

In certain embodiments, the subject treated has no history of myocardialinfarction within three months of treatment according to the methodsprovided herein. In some embodiments, the subject has no history ofcerebrovascular accident or transient ischemic attack within threemonths of treatment according to the methods provided herein. In someembodiments, the subject has no suffered no thromboembelic event,including deep vein thrombosis or pulmonary embolus, within 28 days oftreatment according to the methods provided herein. In otherembodiments, the subject has not experienced or is not experiencinguncontrolled disseminated intravascular coagulation.

Because subjects with cancer have heterogeneous clinical manifestationsand varying clinical outcomes, the treatment given to a patient mayvary, depending on his/her prognosis. The skilled clinician will be ableto readily determine without undue experimentation specific secondaryagents, types of surgery, and types of non-drug based standard therapythat can be effectively used to treat an individual subject with cancer.

It will be appreciated that every suitable combination of the compoundsprovided herein with one or more of the aforementioned compounds andoptionally one or more further pharmacologically active substances iscontemplated herein.

6.5. Formulation of Pharmaceutical Compositions

The pharmaceutical compositions provided herein contain therapeuticallyeffective amounts of one or more solid forms of Compound 1 providedherein and a pharmaceutically acceptable carrier, diluent or excipient.

In one embodiment, the pharmaceutical compositions provided hereincomprise Form A of Compound 1 and one or more pharmaceuticallyacceptable excipients or carriers. In one embodiment, the pharmaceuticalcompositions provided herein comprise Form B of Compound 1 and one ormore pharmaceutically acceptable excipients or carriers. In oneembodiment, the pharmaceutical compositions provided herein compriseForm C of Compound 1 and one or more pharmaceutically acceptableexcipients or carriers. In one embodiment, the pharmaceuticalcompositions provided herein comprise Form D of Compound 1 and one ormore pharmaceutically acceptable excipients or carriers. In oneembodiment, the pharmaceutical compositions provided herein compriseForm E of Compound 1 and one or more pharmaceutically acceptableexcipients or carriers. In one embodiment, the pharmaceuticalcompositions provided herein comprise amorphous Compound 1 and one ormore pharmaceutically acceptable excipients or carriers.

The compounds can be formulated into suitable pharmaceuticalpreparations such as in sterile solutions or suspensions for ophthalmicor parenteral administration, as well as transdermal patch preparationand dry powder inhalers. Typically the solid forms of Compound 1described above are formulated into pharmaceutical compositions usingtechniques and procedures well known in the art (see, e.g., AnselIntroduction to Pharmaceutical Dosage Forms, Seventh Edition 1999).

In the compositions, effective concentrations of one or more solid formsof Compound 1 is mixed with a suitable pharmaceutical carrier orvehicle. In certain embodiments, the concentrations of the solid formsof Compound 1 in the compositions are effective for delivery of anamount, upon administration, that treats, prevents, or ameliorates oneor more of the symptoms and/or progression of cancer, including solidtumors and blood borne tumors.

Typically, the compositions are formulated for single dosageadministration. To formulate a composition, the weight fraction of asolid form of Compound 1 is dissolved, suspended, dispersed or otherwisemixed in a selected vehicle at an effective concentration such that thetreated condition is relieved or ameliorated. Pharmaceutical carriers orvehicles suitable for administration of the solid forms of Compound 1provided herein include any such carriers known to those skilled in theart to be suitable for the particular mode of administration.

In addition, the solid forms of Compound 1 may be formulated as the solepharmaceutically active ingredient in the composition or may be combinedwith other active ingredients. Liposomal suspensions, includingtissue-targeted liposomes, such as tumor-targeted liposomes, may also besuitable as pharmaceutically acceptable carriers. These may be preparedaccording to methods known to those skilled in the art. For example,liposome formulations may be prepared as known in the art. Briefly,liposomes such as multilamellar vesicles (MLV's) may be formed by dryingdown egg phosphatidyl choline and brain phosphatidyl serine (7:3 molarratio) on the inside of a flask. A solution of a compound providedherein in phosphate buffered saline lacking divalent cations (PBS) isadded and the flask shaken until the lipid film is dispersed. Theresulting vesicles are washed to remove unencapsulated compound,pelleted by centrifugation, and then resuspended in PBS.

A solid form of Compound 1 is included in the pharmaceuticallyacceptable carrier in an amount sufficient to exert a therapeuticallyuseful effect in the absence of undesirable side effects on the patienttreated. The therapeutically effective concentration may be determinedempirically by testing the solid forms of Compound 1 in in vitro and invivo systems described herein and then extrapolated therefrom fordosages for humans.

The concentration of a solid form of Compound 1 in the pharmaceuticalcomposition will depend on absorption, tissue distribution, inactivationand excretion rates of the solid form of Compound 1, the physicochemicalcharacteristics of the solid form of Compound 1, the dosage schedule,and amount administered as well as other factors known to those of skillin the art. For example, the amount that is delivered is sufficient toameliorate one or more of the symptoms of cancer, including solid tumorsand blood borne tumors.

In certain embodiments, a therapeutically effective dosage shouldproduce a serum concentration of active ingredient of from about 0.1ng/mL to about 50-100 μg/mL. In one embodiment, the pharmaceuticalcompositions provide a dosage of from about 0.001 mg to about 2000 mg ofcompound per kilogram of body weight per day. Pharmaceutical dosage unitforms are prepared to provide from about 1 mg to about 1000 mg and incertain embodiments, from about 10 to about 500 mg of the essentialactive ingredient or a combination of essential ingredients per dosageunit form.

The active ingredient may be administered at once, or may be dividedinto a number of smaller doses to be administered at intervals of time.It is understood that the precise dosage and duration of treatment is afunction of the disease being treated and may be determined empiricallyusing known testing protocols or by extrapolation from in vivo or invitro test data. It is to be noted that concentrations and dosage valuesmay also vary with the severity of the condition to be alleviated. It isto be further understood that for any particular subject, specificdosage regimens should be adjusted over time according to the individualneed and the professional judgment of the person administering orsupervising the administration of the compositions, and that theconcentration ranges set forth herein are exemplary only and are notintended to limit the scope or practice of the claimed compositions.

Thus, effective concentrations or amounts of one or more of the solidforms of Compound 1 described herein are mixed with a suitablepharmaceutical carrier or vehicle for systemic, topical or localadministration to form pharmaceutical compositions. The solid forms ofCompound 1 are included in an amount effective for ameliorating one ormore symptoms of, or for treating, retarding progression, or preventing.The concentration of a solid form of Compound 1 in the composition willdepend on absorption, tissue distribution, inactivation, excretion ratesof the solid form of Compound 1, the dosage schedule, amountadministered, particular formulation as well as other factors known tothose of skill in the art.

The compositions are intended to be administered by a suitable route,including but not limited to parenterally, rectally, topically andlocally. The compositions are in liquid, semi-liquid or solid form andare formulated in a manner suitable for each route of administration.

Solutions or suspensions used for parenteral, intradermal, subcutaneous,or topical application can include any of the following components: asterile diluent, such as water for injection, saline solution, fixedoil, polyethylene glycol, glycerine, propylene glycol, dimethylacetamide or other synthetic solvent; antimicrobial agents, such asbenzyl alcohol and methyl parabens; antioxidants, such as ascorbic acidand sodium bisulfite; chelating agents, such asethylenediaminetetraacetic acid (EDTA); buffers, such as acetates,citrates and phosphates; and agents for the adjustment of tonicity suchas sodium chloride or dextrose. Parenteral preparations can be enclosedin ampules, pens, disposable syringes or single or multiple dose vialsmade of glass, plastic or other suitable material.

In instances in which the solid forms of Compound 1 exhibit insufficientsolubility, methods for solubilizing compounds may be used. Such methodsare known to those of skill in this art, and include, but are notlimited to, using cosolvents, such as dimethylsulfoxide (DMSO), usingsurfactants, such as TWEEN®, or dissolution in aqueous sodiumbicarbonate.

Upon mixing or addition of the compound(s), the resulting mixture may bea solution, suspension, emulsion or the like. The form of the resultingmixture depends upon a number of factors, including the intended mode ofadministration and the solubility of the solid form of Compound 1 in theselected carrier or vehicle. The effective concentration is sufficientfor ameliorating the symptoms of the disease, disorder or conditiontreated and may be empirically determined.

The pharmaceutical compositions are provided for administration tohumans and animals in unit dosage forms, such as powders, sterileparenteral solutions or suspensions, and oil water emulsions containingsuitable quantities of the solid forms of Compound 1. The solid forms ofCompound 1 are formulated and administered in unit dosage forms ormultiple dosage forms. Unit dose forms as used herein refer tophysically discrete units suitable for human and animal subjects andpackaged individually as is known in the art. Each unit dose contains apredetermined quantity of the solid form of Compound 1 sufficient toproduce the desired therapeutic effect, in association with the requiredpharmaceutical carrier, vehicle or diluent. Examples of unit dose formsinclude ampules and syringes. Unit dose forms may be administered infractions or multiples thereof. A multiple dose form is a plurality ofidentical unit dosage forms packaged in a single container to beadministered in segregated unit dose form. Examples of multiple doseforms include vials, or bottles of pints or gallons. Hence, multipledose form is a multiple of unit doses which are not segregated inpackaging.

Sustained-release preparations can also be prepared. Suitable examplesof sustained-release preparations include semipermeable matrices ofsolid hydrophobic polymers containing a solid form of Compound 1provided herein, which matrices are in the form of shaped articles,e.g., films, or microcapsule. Examples of sustained-release matricesinclude iontophoresis patches, polyesters, hydrogels (for example,poly(2-hydroxyethyl-methacrylate), or poly(vinylalcohol)), polylactides,copolymers of L-glutamic acid and ethyl-L-glutamate, non-degradableethylene-vinyl acetate, degradable lactic acid-glycolic acid copolymerssuch as the LUPRON DEPOT™ (injectable microspheres composed of lacticacid-glycolic acid copolymer and leuprolide acetate), andpoly-D-(−)-3-hydroxybutyric acid. While polymers such as ethylene-vinylacetate and lactic acid-glycolic acid enable release of molecules forover 100 days, certain hydrogels release proteins for shorter timeperiods. When encapsulated compound remain in the body for a long time,they may denature or aggregate as a result of exposure to moisture at37° C., resulting in a loss of biological activity and possible changesin their structure. Rational strategies can be devised for stabilizationdepending on the mechanism of action involved. For example, if theaggregation mechanism is discovered to be intermolecular S—S bondformation through thio-disulfide interchange, stabilization may beachieved by modifying sulfhydryl residues, lyophilizing from acidicsolutions, controlling moisture content, using appropriate additives,and developing specific polymer matrix compositions.

Dosage forms or compositions containing a solid form of Compound 1 inthe range of 0.005% to 100% with the balance made up from non toxiccarrier may be prepared.

The compositions may include other active compounds to obtain desiredcombinations of properties. The solid forms of Compound 1 providedherein may also be advantageously administered for therapeutic orprophylactic purposes together with another pharmacological agent knownin the general art to be of value in treating one or more of thediseases or medical conditions referred to hereinabove, such as diseasesrelated to oxidative stress. It is to be understood that suchcombination therapy constitutes a further aspect of the compositions andmethods of treatment provided herein.

Further encompassed are anhydrous pharmaceutical compositions and dosageforms containing a solid form of Compound 1 provided herein. Forexample, the addition of water (e.g., 5%) is widely accepted in thepharmaceutical arts as a means of simulating long-term storage in orderto determine characteristics such as shelf-life or the stability offormulations over time. See, e.g., Jens T. Carstensen, Drug Stability:Principles & Practice, 2 d. Ed., Marcel Dekker, NY, N.Y., 1995, pp.379-80. In effect, water and heat accelerate the decomposition of somecompounds. Thus, the effect of water on a formulation can be of greatsignificance since moisture and/or humidity are commonly encounteredduring manufacture, handling, packaging, storage, shipment and use offormulations.

Anhydrous pharmaceutical compositions and dosage forms provided hereincan be prepared using anhydrous or low moisture containing ingredientsand low moisture or low humidity conditions. Pharmaceutical compositionsand dosage forms that comprise lactose and at least one activeingredient that comprises a primary or secondary amine are anhydrous ifsubstantial contact with moisture and/or humidity during manufacturing,packaging, and/or storage is expected.

An anhydrous pharmaceutical composition should be prepared and storedsuch that its anhydrous nature is maintained. Accordingly, anhydrouscompositions are packaged using materials known to prevent exposure towater such that they can be included in suitable formulary kits.Examples of suitable packaging include, but are not limited to,hermetically sealed foils, plastics, unit dose containers (e.g., vials).

6.5.1. Injectables, Solutions and Emulsions

Parenteral administration, generally characterized by injection, eithersubcutaneously, intramuscularly or intravenously is also contemplatedherein. Injectables can be prepared in conventional forms, either asliquid solutions or suspensions, solid forms suitable for solution orsuspension in liquid prior to injection, or as emulsions. Suitableexcipients are, for example, water, saline, dextrose, glycerol orethanol. In addition, if desired, the pharmaceutical compositions to beadministered may also contain minor amounts of non toxic auxiliarysubstances such as wetting or emulsifying agents, pH buffering agents,stabilizers, solubility enhancers, and other such agents, such as forexample, sodium acetate, sorbitan monolaurate, triethanolamine oleateand cyclodextrins. Implantation of a slow release or sustained releasesystem, such that a constant level of dosage is maintained is alsocontemplated herein. Briefly, a solid form of Compound 1 provided hereinis dispersed in a solid inner matrix, e.g., polymethylmethacrylate,polybutylmethacrylate, plasticized or unplasticized polyvinylchloride,plasticized nylon, plasticized polyethyleneterephthalate, naturalrubber, polyisoprene, polyisobutylene, polybutadiene, polyethylene,ethylene-vinylacetate copolymers, silicone rubbers,polydimethylsiloxanes, silicone carbonate copolymers, hydrophilicpolymers such as hydrogels of esters of acrylic and methacrylic acid,collagen, cross-linked polyvinylalcohol and cross-linked partiallyhydrolyzed polyvinyl acetate, that is surrounded by an outer polymericmembrane, e.g., polyethylene, polypropylene, ethylene/propylenecopolymers, ethylene/ethyl acrylate copolymers, ethylene/vinylacetatecopolymers, silicone rubbers, polydimethyl siloxanes, neoprene rubber,chlorinated polyethylene, polyvinylchloride, vinylchloride copolymerswith vinyl acetate, vinylidene chloride, ethylene and propylene, ionomerpolyethylene terephthalate, butyl rubber epichlorohydrin rubbers,ethylene/vinyl alcohol copolymer, ethylene/vinyl acetate/vinyl alcoholterpolymer, and ethylene/vinyloxyethanol copolymer, that is insoluble inbody fluids. The compound diffuses through the outer polymeric membranein a release rate controlling step. The percentage of active compoundcontained in such parenteral compositions is highly dependent on thespecific nature thereof, as well as the activity of the compound and theneeds of the subject.

Parenteral administration of the compositions includes intravenous,subcutaneous and intramuscular administrations. Preparations forparenteral administration include sterile solutions ready for injection,sterile dry soluble products, such as lyophilized powders, ready to becombined with a solvent just prior to use, including hypodermic tablets,sterile suspensions ready for injection, sterile dry insoluble productsready to be combined with a vehicle just prior to use and sterileemulsions. The solutions may be either aqueous or nonaqueous.

If administered intravenously, suitable carriers include physiologicalsaline or phosphate buffered saline (PBS), and solutions containingthickening and solubilizing agents, such as glucose, polyethyleneglycol, and polypropylene glycol and mixtures thereof.

Pharmaceutically acceptable carriers used in parenteral preparationsinclude aqueous vehicles, nonaqueous vehicles, antimicrobial agents,isotonic agents, buffers, antioxidants, local anesthetics, suspendingand dispersing agents, emulsifying agents, sequestering or chelatingagents and other pharmaceutically acceptable substances.

Examples of aqueous vehicles include Sodium Chloride Injection, RingersInjection, Isotonic Dextrose Injection, Sterile Water Injection,Dextrose and Lactated Ringers Injection. Nonaqueous parenteral vehiclesinclude fixed oils of vegetable origin, cottonseed oil, corn oil, sesameoil and peanut oil. Antimicrobial agents in bacteriostatic orfungistatic concentrations must be added to parenteral preparationspackaged in multiple dose containers which include phenols or cresols,mercurials, benzyl alcohol, chlorobutanol, methyl and propyl phydroxybenzoic acid esters, thimerosal, benzalkonium chloride andbenzethonium chloride. Isotonic agents include sodium chloride anddextrose. Buffers include phosphate and citrate. Antioxidants includesodium bisulfate. Local anesthetics include procaine hydrochloride.Suspending and dispersing agents include sodium carboxymethylcelluose,hydroxypropyl methylcellulose and polyvinylpyrrolidone. Emulsifyingagents include Polysorbate 80 (TWEEN® 80). A sequestering or chelatingagent of metal ions include EDTA. Pharmaceutical carriers also includeethyl alcohol, polyethylene glycol and propylene glycol for watermiscible vehicles and sodium hydroxide, hydrochloric acid, citric acidor lactic acid for pH adjustment.

The concentration of the pharmaceutically active compound is adjusted sothat an injection provides an effective amount to produce the desiredpharmacological effect. The exact dose depends on the age, weight andcondition of the patient or animal as is known in the art.

The unit dose parenteral preparations are packaged in an ampule, a vialor a syringe with a needle. All preparations for parenteraladministration must be sterile, as is known and practiced in the art.

Illustratively, intravenous or intraarterial infusion of a sterileaqueous solution containing a solid form of Compound 1 is an effectivemode of administration. Another embodiment is a sterile aqueous or oilysolution or suspension containing an active material injected asnecessary to produce the desired pharmacological effect.

Injectables are designed for local and systemic administration.Typically a therapeutically effective dosage is formulated to contain aconcentration of at least about 0.1% w/w up to about 90% w/w or more,such as more than 1% w/w of a solid forms of Compound 1 to the treatedtissue(s). The active ingredient may be administered at once, or may bedivided into a number of smaller doses to be administered at intervalsof time. It is understood that the precise dosage and duration oftreatment is a function of the tissue being treated and may bedetermined empirically using known testing protocols or by extrapolationfrom in vivo or in vitro test data. It is to be noted thatconcentrations and dosage values may also vary with the age of theindividual treated. It is to be further understood that for anyparticular subject, specific dosage regimens should be adjusted overtime according to the individual need and the professional judgment ofthe person administering or supervising the administration of theformulations, and that the concentration ranges set forth herein areexemplary only and are not intended to limit the scope or practice ofthe claimed formulations.

A solid form of Compound 1 may be suspended in micronized or othersuitable form or may be derivatized to produce a more soluble activeproduct or to produce a prodrug. The form of the resulting mixturedepends upon a number of factors, including the intended mode ofadministration and the solubility of the solid form of Compound 1 in theselected carrier or vehicle. The effective concentration is sufficientfor ameliorating the symptoms of the condition and may be empiricallydetermined.

6.5.2. Lyophilized Powders

Of interest herein are also lyophilized powders, which can bereconstituted for administration as solutions, emulsions and othermixtures. They may also be reconstituted and formulated as solids orgels.

The sterile, lyophilized powder is prepared by dissolving a solid formof Compound 1 provided herein in a suitable solvent. The solvent maycontain an excipient which improves the stability or otherpharmacological component of the powder or reconstituted solution,prepared from the powder. Excipients that may be used include, but arenot limited to, dextrose, sorbital, fructose, corn syrup, xylitol,glycerin, glucose, sucrose or other suitable agent. The solvent may alsocontain a buffer, such as citrate, sodium or potassium phosphate orother such buffer known to those of skill in the art at, in oneembodiment, about neutral pH. Subsequent sterile filtration of thesolution followed by lyophilization under standard conditions known tothose of skill in the art provides the desired formulation. Generally,the resulting solution will be apportioned into vials forlyophilization. Each vial will contain a single dosage (including butnot limited to 1-1000 mg or 100-500 mg) or multiple dosages of thecompound. The lyophilized powder can be stored under appropriateconditions, such as at about 4° C. to room temperature.

Reconstitution of this lyophilized powder with water for injectionprovides a formulation for use in parenteral administration. Forreconstitution, about 0.1-50 mg, about 0.5-50 mg, about 1 to 50 mg,about 5-35 mg, or about 9-30 mg of lyophilized powder, is added per mLof sterile water or other suitable carrier. In one embodiment, about 0.5mg of lyophilized powder is added per mL of sterile water or othersuitable carrier.

Exemplary lyophilized formulations comprising the solid forms providedherein (e.g., Form A, Form B, Form C, Form D, Form E and/or amorphous ofCompound 1) are described in a U.S. provisional patent application filedconcurrently herewith entitled “FORMULATIONS OF3-(5-AMINO-2-METHYL-4-OXO-4H-QUINAZOLIN-3-YL)-PIPERIDINE-2,6-DIONE”, thedisclosure of which is incorporated herein by reference in its entirety.

6.5.3. Topical Administration

Topical mixtures are prepared as described for the local and systemicadministration. The resulting mixture may be a solution, suspension,emulsion or the like and are formulated as creams, gels, ointments,emulsions, solutions, elixirs, lotions, suspensions, tinctures, pastes,foams, aerosols, irrigations, sprays, suppositories, bandages, dermalpatches or any other formulations suitable for topical administration.

The solid forms of Compound 1 thereof may be formulated as aerosols fortopical application, such as by inhalation (see, e.g., U.S. Pat. Nos.4,044,126, 4,414,209, and 4,364,923, which describe aerosols fordelivery of a steroid useful for treatment of inflammatory diseases,particularly asthma). These formulations for administration to therespiratory tract can be in the form of an aerosol or solution for anebulizer, or as a microfine powder for insufflation, alone or incombination with an inert carrier such as lactose. In such a case, theparticles of the formulation will have diameters of less than 50 micronsor less than 10 microns.

The solid forms of Compound 1 may be formulated for local or topicalapplication, such as for topical application to the skin and mucousmembranes, such as in the eye, in the form of gels, creams, and lotionsand for application to the eye or for intracisternal or intraspinalapplication. Topical administration is contemplated for transdermaldelivery and also for administration to the eyes or mucosa, or forinhalation therapies. Nasal solutions of the solid forms of Compound 1alone or in combination with other pharmaceutically acceptableexcipients can also be administered.

These solutions, particularly those intended for ophthalmic use, may beformulated as 0.01%-10% isotonic solutions, pH about 5-7, withappropriate salts.

6.5.4. Compositions for Other Routes of Administration

Other routes of administration, such as topical application, transdermalpatches, and rectal administration are also contemplated herein.

For example, pharmaceutical dosage forms for rectal administration arerectal suppositories, capsules and tablets for systemic effect. Rectalsuppositories are used herein mean solid bodies for insertion into therectum which melt or soften at body temperature releasing one or morepharmacologically or therapeutically active ingredients.Pharmaceutically acceptable substances utilized in rectal suppositoriesare bases or vehicles and agents to raise the melting point. Examples ofbases include cocoa butter (theobroma oil), glycerin gelatin, carbowax(polyoxyethylene glycol) and appropriate mixtures of mono, di andtriglycerides of fatty acids. Combinations of the various bases may beused. Agents to raise the melting point of suppositories includespermaceti and wax. Rectal suppositories may be prepared either by thecompressed method or by molding. An exemplary weight of a rectalsuppository is about 2 to 3 grams.

6.5.5. Sustained Release Compositions

Active ingredients provided herein can be administered by controlledrelease means or by delivery devices that are well known to those ofordinary skill in the art. Examples include, but are not limited to,those described in U.S. Pat. Nos. 3,845,770; 3,916,899; 3,536,809;3,598,123; and 4,008,719, 5,674,533, 5,059,595, 5,591,767, 5,120,548,5,073,543, 5,639,476, 5,354,556, 5,639,480, 5,733,566, 5,739,108,5,891,474, 5,922,356, 5,972,891, 5,980,945, 5,993,855, 6,045,830,6,087,324, 6,113,943, 6,197,350, 6,248,363, 6,264,970, 6,267,981,6,376,461, 6,419,961, 6,589,548, 6,613,358, 6,699,500 and 6,740,634,each of which is incorporated herein by reference. Such dosage forms canbe used to provide slow or controlled-release of one or more activeingredients using, for example, hydropropylmethyl cellulose, otherpolymer matrices, gels, permeable membranes, osmotic systems, multilayercoatings, microparticles, liposomes, microspheres, or a combinationthereof to provide the desired release profile in varying proportions.Suitable controlled-release formulations known to those of ordinaryskill in the art, including those described herein, can be readilyselected for use with the active ingredients provided herein.

All controlled-release pharmaceutical products have a common goal ofimproving drug therapy over that achieved by their non-controlledcounterparts. In one embodiment, the use of an optimally designedcontrolled-release preparation in medical treatment is characterized bya minimum of drug substance being employed to cure or control thecondition in a minimum amount of time. In certain embodiments,advantages of controlled-release formulations include extended activityof the drug, reduced dosage frequency, and increased patient compliance.In addition, controlled-release formulations can be used to affect thetime of onset of action or other characteristics, such as blood levelsof the drug, and can thus affect the occurrence of side (e.g., adverse)effects.

Most controlled-release formulations are designed to initially releasean amount of drug (active ingredient) that promptly produces the desiredtherapeutic effect, and gradually and continually release of otheramounts of drug to maintain this level of therapeutic or prophylacticeffect over an extended period of time. In order to maintain thisconstant level of drug in the body, the drug must be released from thedosage form at a rate that will replace the amount of drug beingmetabolized and excreted from the body. Controlled-release of an activeingredient can be stimulated by various conditions including, but notlimited to, pH, temperature, enzymes, water, or other physiologicalconditions or solid forms of Compound 1.

In certain embodiments, the agent may be administered using intravenousinfusion, an implantable osmotic pump, a transdermal patch, liposomes,or other modes of administration. In one embodiment, a pump may be used(see, Sefton, CRC Crit. Ref. Biomed. Eng. 14:201 (1987); Buchwald etal., Surgery 88:507 (1980); Saudek et al., N. Engl. J. Med. 321:574(1989). In another embodiment, polymeric materials can be used. In yetanother embodiment, a controlled release system can be placed inproximity of the therapeutic target, i.e., thus requiring only afraction of the systemic dose (see, e.g., Goodson, Medical Applicationsof Controlled Release, vol. 2, pp. 115-138 (1984).

In some embodiments, a controlled release device is introduced into asubject in proximity of the site of inappropriate immune activation or atumor. Other controlled release systems are discussed in the review byLanger (Science 249:1527-1533 (1990). The active ingredient can bedispersed in a solid inner matrix, e.g., polymethylmethacrylate,polybutylmethacrylate, plasticized or unplasticized polyvinylchloride,plasticized nylon, plasticized polyethyleneterephthalate, naturalrubber, polyisoprene, polyisobutylene, polybutadiene, polyethylene,ethylene-vinylacetate copolymers, silicone rubbers,polydimethylsiloxanes, silicone carbonate copolymers, hydrophilicpolymers such as hydrogels of esters of acrylic and methacrylic acid,collagen, cross-linked polyvinylalcohol and cross-linked partiallyhydrolyzed polyvinyl acetate, that is surrounded by an outer polymericmembrane, e.g., polyethylene, polypropylene, ethylene/propylenecopolymers, ethylene/ethyl acrylate copolymers, ethylene/vinylacetatecopolymers, silicone rubbers, polydimethyl siloxanes, neoprene rubber,chlorinated polyethylene, polyvinylchloride, vinylchloride copolymerswith vinyl acetate, vinylidene chloride, ethylene and propylene, ionomerpolyethylene terephthalate, butyl rubber epichlorohydrin rubbers,ethylene/vinyl alcohol copolymer, ethylene/vinyl acetate/vinyl alcoholterpolymer, and ethylene/vinyloxyethanol copolymer, that is insoluble inbody fluids. The active ingredient then diffuses through the outerpolymeric membrane in a release rate controlling step. The percentage ofactive ingredient contained in such parenteral compositions is highlydependent on the specific nature thereof, as well as the needs of thesubject.

6.5.6. Targeted Formulations

The solid forms of Compound 1 provided herein may also be formulated tobe targeted to a particular tissue, receptor, or other area of the bodyof the subject to be treated. Many such targeting methods are well knownto those of skill in the art. All such targeting methods arecontemplated herein for use in the instant compositions. Fornon-limiting examples of targeting methods, see, e.g., U.S. Pat. Nos.6,316,652, 6,274,552, 6,271,359, 6,253,872, 6,139,865, 6,131,570,6,120,751, 6,071,495, 6,060,082, 6,048,736, 6,039,975, 6,004,534,5,985,307, 5,972,366, 5,900,252, 5,840,674, 5,759,542 and 5,709,874.

In one embodiment, liposomal suspensions, including tissue-targetedliposomes, such as tumor-targeted liposomes, may also be suitable aspharmaceutically acceptable carriers. These may be prepared according tomethods known to those skilled in the art. For example, liposomeformulations may be prepared as described in U.S. Pat. No. 4,522,811.Briefly, liposomes such as multilamellar vesicles (MLV's) may be formedby drying down egg phosphatidyl choline and brain phosphatidyl serine(7:3 molar ratio) on the inside of a flask. A solution of a solid formof Compound 1 provided herein in phosphate buffered saline lackingdivalent cations (PBS) is added and the flask shaken until the lipidfilm is dispersed. The resulting vesicles are washed to removeunencapsulated compound, pelleted by centrifugation, and thenresuspended in PBS.

6.5.7. Articles of Manufacture

The solid forms of Compound 1 provided herein which is used fortreatment, prevention or amelioration of one or more symptoms orprogression of cancer, including solid tumors and blood borne tumors,and a label that indicates that the solid form of Compound 1 is used fortreatment, prevention or amelioration of one or more symptoms orprogression of cancer, including solid tumors and blood borne tumors.

The articles of manufacture provided herein contain packaging materials.Packaging materials for use in packaging pharmaceutical products arewell known to those of skill in the art. See, e.g., U.S. Pat. Nos.5,323,907, 5,052,558 and 5,033,252. Examples of pharmaceutical packagingmaterials include, but are not limited to, blister packs, bottles,tubes, inhalers, pumps, bags, vials, containers, syringes, pens,bottles, and any packaging material suitable for a selected formulationand intended mode of administration and treatment. A wide array offormulations of the compounds and compositions provided herein arecontemplated.

6.6. Evaluation of Activity

Standard physiological, pharmacological and biochemical procedures areavailable for testing the compounds to identify those that possess thedesired anti-proliferative activity.

Such assays include, for example, biochemical assays such as bindingassays, radioactivity incorporation assays, as well as a variety of cellbased assays, including KG-1 cell proliferation assay described in theExample section.

7. Examples

The following Examples are presented by way of illustration, notlimitation. The following abbreviations are used in descriptions andexamples:

BuOH Butanol

DCM: Dichloromethane

DMSO: Dimethylsulfoxide

DSC: Differential Scanning calorimetry

DVS Dynamic vapor sorption

EDTA: Ethylenediamine tetraacetate

EtOH: Ethanol

EtOAc Ethyl acetate

HPLC: High performance liquid chromatography

IPA: 2-Propanol

KF Karl Fisher

LCMS: Liquid Chromatography with Mass Spectroscopy

MeCN Acetonitrile

MeOAc: Methylacetate

MeOH: Methanol

MEK: Methyl ethyl ketone

MIBK: Methyl isobutyl ketone

MTBE Methyl t-butyl ether

NMP: N-Methyl-2-pyrrolidone

NMR: Nuclear magnetic resonance

RH: Relative Humidity

RT: Room Temperature

SEM Scanning Electron Microscope °

TA: Thermal Analysis

TGA: Thermogravimetric Analysis

TGA-MS/TG-MS: Thermogravimetric Analysis coupled with Mass Spectroscopy

THF: Tetrahydrofuran

TLC: Thin layer chromatography

XRPD: X-Ray Powder Diffraction

7.1 Solid Forms 7.1.1 Polymorph Screen

A polymorph screen of Compound 1 was performed to investigate whetherdifferent solid forms could be generated under various conditions, suchas different solvents, temperature and humidity changes.

The solvents used in the polymorph screen were either HPLC or reagentgrade, including acetone, acetonitrile (MeCN), MeCN/water (1:1),n-butanol (n-BuOH), absolute ethanol (EtOH), ethanol/water (1:1),methanol (MeOH), 2-propanol (IPA), ethyl acetate (EtOAc), methyl acetate(MeOAc), dichloromethane (DCM), methyl ethyl ketone (MEK), methylt-butyl ether (MTBE), heptane, toluene, tetrahydrofuran (THF), THF/water(1:1), water, dimethyl sufoxide (DMSO), and N-methylpyrrolidone (NMP).

The characterization of the crystal forms produced during the screen wasperformed by X-ray powder diffraction (XRPD), differential scanningcalorimetry (DSC), thermogravimetric analysis (TGA), Miniature ScanningElectron Microscope (Mini SEM) and dynamic vapor sorption (DVS), KarlFischer (KF), and/or ¹H-Nuclear Magnetic Resonance (NMR).

XRPD analysis was conducted on a PANalytical Empyrean X-ray powderdiffractometer using Cu Kα radiation at 1.54 Å. The PANalytical Empyreaninstrument was equipped with a fine focus X-ray tube. The voltage andamperage of the X-ray generator were set at 45 kV and 40 mA,respectively. The divergence slits were set at 1/16° and ⅛°, and thereceiving slits was set at 1/16°. Diffracted radiation was measuredusing a Pixel 2D detector. A theta-two theta continuous scan was set atstep size 0.013 or 0.026 from 3° to 40° 2θ with sample spinning rate at4. A sintered alumina standard was used to check the peak positions.

XRPD data were obtained on a PANalytical Empyrean X-ray powderdiffractometer using similar instrument parameters.

DSC analyses were performed on a TA Discovery Differential Scanningcalorimeter. Indium was used as the calibration standard. Approximately1-5 mg of sample was placed into a DSC pan. The sample was heated undernitrogen at a rate of 10° C./min, up to a final temperature of 300° C.Melting points were reported as the extrapolated onset temperatures.

TGA analyses were performed on a TA Discovery ThermogravimetricAnalyzer. Calcium oxalate was used for a performance check.Approximately 2-10 mg of accurately weighed sample was placed on a panand loaded into the TGA furnace. The sample was heated under nitrogen ata rate of 10° C./min, up to a final temperature of 300° C.

TGA/SDTA analyses were performed on a TGA/SDTA851e instrument(Mettler-Toledo GmbH, Switzerland). The TGA/SDTA851e instrument wascalibrated for temperature with indium and aluminium. Samples wereweighed into 100 μl aluminium crucibles and sealed. The seals werepin-holed and the crucibles heated in the TGA from 25 to 300° C. at aheating rate of 10° C./min. Dry N2 gas was used for purging. The TGAdata were obtained on a TA Q500 TGA instrument. The samples were heatedat a rate of 10° C./min using nitrogen as the purge gas.

Morphology analysis of the samples was carried out on an Evex Mini SEM.Small amounts of samples were dispersed on a sample holder, and thencoating with gold and viewed with 500× to 1000× magnification.

Hygroscopicity was determined on a Surface Measurement Systems DVS.Typically a sample size of 5-20 mg was loaded into the DVS instrumentsample pan and the sample was analyzed on a DVS automated sorptionanalyzer at room temperature. The relative humidity was increased from0% to 90% RH at 10% RH step, then decreased in a similar manner toaccomplish a full adsorption/desorption cycle.

¹H NMR spectra were obtained on a Bruker 300 MHz NMR spectrometer.Samples were dissolved in DMSO-d6 and analyzed with 8-64 scans.

Solubility of Form C in selected aqueous and organic solvents wasdetermined by mixing solid with solvents at room temperature. Thesolubility samples were filtered after 24 hr of agitation and quantifiedby an HPLC method, except for MeOH in which the solubility wasdetermined after agitating 2 hr in order to minimize degradation.

Water content was measured using a coulometric KF titrator equipped withan oven sample processor. The oven temperature was set as 225° C.

7.1.2 Experiments and Methods 7.1.2.1 Solubility

In order to select the screening solvents and to determine theconcentration range to be used in the screen, a quantitative solubilityassessment was performed on Form B and Form C of Compound 1. Approximatesolubility of Compound 1 Form B and solubility of Form C by HPLC at roomtemperature are provided in Tables 1 and 2, respectively.

TABLE 1 Approximate Solubility of Compound 1 Form B at Room Temperature.Approximate Solubility Solvent (mg/mL) Acetone <6 MeCN <3 MeCN/water(1:1) <3 n-BuOH <2 EtOH <2 EtOH/water (1:1) <2 MeOH <15* IPA <1 EtOAc <2MEK <3 DCM <1 MTBE <1 Heptane <1 Toluene <1 THF <12  THF/water (1:1)<10  Water <1 MeOAc <3 NMP >100  DMSO >100  *degradation suspected.

TABLE 2 HPLC Solubility of Form C in Selected Solvents at RoomTemperature. Solubility Solvent (mg/mL) Water 0.001 0.9% NaCl 0.001 0.1NHCl 0.001 Acetate buffer pH 4.0 0.001 Phosphate buffer pH 6.8 0.001 MeCN1.179 Acetone 2.354 MeOH 0.843 EtOH 0.235 IPA 0.071 EtOAc 0.285 THF4.349 Heptane <0.0001 Toluene 0.002 DMSO >200

7.1.2.2 Equilibration and Evaporation

Equilibration and evaporation experiments were performed at roomtemperature and 50° C. using Compound 1 Form B as starting material. Anexcess of Compound 1 Form B was added to up to 2 mL of a test solvent.The resulting mixture was agitated for 4 days at room temperature andfor about 18 hours at 50° C. separately. Upon reaching equilibrium, thesaturated supernatant solution was removed, filtered using 0.45 μm PTFEfilters and allowed to evaporate in an open vial under nitrogen at roomtemperature and 50° C., respectively. The solid resulting from theequilibration was isolated and air-dried before analysis. The resultsare summarized in Table 3.

TABLE 3 Summary of Equilibration and Evaporation Results. Form by XRPDSolvent EQ at RT EV at RT EQ at 50° C. EV at 50° C. Acetone B — A — MeCNB — A — MeCN/water B — C — n-BuOH B — B (+A) — EtOH B — A — EtOH/water B— C — MeOH B + A amorphous* — amorphous* IPA B — B + A — EtOAc B — B + A— MEK B + A — A — DCM B — n/a — MTBE B + A — n/a — Heptane B — B —Toluene B — B — THF B + A B A + B B THF/water B + A B B + A B water B —B — MeOAc B — A — n/a: no experiment —: not analyzable. *significantdegradation occurred.

Equilibration in MeCN/water and EtOH/water at 50° C. afforded Compound 1Form C. Equilibration in EtOH, MEK, and MeOAc at 50° C. afforded Form A.All other equilibration experiments afforded Form B or Form B mixed withForm A or Form C. Due to relatively low solubility, most evaporationexperiments didn't afford analyzable solid. Evaporation from THF andTHF/water afforded Form B. Solids from MeOH evaporation showed amorphousXRPD patterns, but ¹H NMR spectrum of the solid showed significantdegradation.

7.1.2.2 Cooling Recrystallization

Cooling recrystallization experiments were performed as described inSection 4.3. The solvents included MeOH/water (1:1), THF/water (1:1),and THF. The results are summarized in Table 4. The solids obtained fromTHF/water was confirmed to be Form B. A small amount of solid wasobtained from MeOH/water, but showed a diffuse XRPD pattern from whichthe solid form can't be identified. Precipitation was not observed fromthe THF recystallization experiment.

TABLE 4 Results from Cooling Recrystallization Solvent Cooling ProfileForm by XRPD MeOH/water (1:1) 70 to −15° C. ? (few peaks) THF/water(1:1) 70 to −15° C. B THF 70 to −15° C. — — no precipitation

7.1.2.3 Recrystallizations with Anti-Solvents

Recrystallizations with anti-solvents were performed as described inSection 4.3. MeOH and DMSO were used as primary solvents. MeCN, acetone,heptane, EtOAc, toluene, water, and IPA were used as anti-solvents. Theresults are summarized in Table 5. Precipitation was observed only froma few solvent systems, including MeOH/water, DMSO/IPA, DMSO/toluene, andDMSO/water, and all solids were confirmed to be Form B.

Primary Anti- Solvent Cooling Form by solvent Solvent Ratio profile XRPDMeOH MeCN 1:15 50 to 4° C. — MeOH acetone 1:15 50 to 4° C. — MeOHheptane 1:15 50 to 4° C. — MeOH EtOAc 1:15 50 to 4° C. — MeOH toluene1:15 50 to 4° C. — MeOH water 1:15 50 to 4° C. B DMSO acetone 1:15 RT to4° C. — DMSO MeCN 1:15 RT to 4° C. — DMSO IPA 1:15 RT to 4° C. B DMSOheptane 1:15 RT to 4° C. — DMSO toluene 1:15 RT to 4° C. B DMSO water1:15 RT to 4° C. B RT: room temperature —: no precipitation

7.1.2.4 Slurry and Recrystallizations

Additional experiments were performed to generated materials for furthercharacterization, as detailed in Table 6.

TABLE 6 Experiments to Generate Materials for Characterization SolventExperimental Conditions Form by XRPD MeCN/water Slurry, shaking at 50°C. for 1 day C (1:1) EtOH/water Slurry, shaking at 50° C. for 1 day A(1:1) EtOH/water Slurry, shaking at 50° C. for 3 days C (1:1) THF/waterCooling recrystallization from ~70° C. B (1:1) to RT (highconcentration) THF/water Cooling recrystallization from ~70° C. B (1:1)to RT (low concentration)

7.1.2.5 Interconversion Among Solid Forms

Further form conversion experiments were performed to determineinterconversion among solid forms. The results are summarized in Table7.

TABLE 7 Summary of Form Transfer Experiments Starting Solvent/Temperature/ Resulting Form(s) Condition Condition Form(s) A + B + Cslurry in acetone RT, 3 days C A + C slurry in EtOH/water RT, 1 day C(1:1) B + unknown slurry in acetone 50° C., 1 day C A/C mix B + unknownslurry in acetone RT, 1 day A A/C mix B + unknown slurry in heptane 50°C., 3 days A + B A/C mix B + unknown slurry in heptane RT, 4 days A + BA/C mix B + unknown slurry in IPA 50° C., 2 days A A/C mix B + unknownslurry in IPA RT, 2 days A + C + A/C mix B? B + unknown slurry inIPA/water RT, 4 days A A/C mix B + unknown slurry in IPA/water RT, 4days A A/C mix (2:1) B + unknown slurry in IPA/water RT, 4 days A A/Cmix (3:1) B + unknown slurry in IPA/water RT, 4 days A A/C mix (5:1) Aheating 225° C., ~10 min in A KF oven B heating 160° C., 1 min B Bheating 225° C., ~10 min in A KF oven C heating 225° C., ~10 min in C KFoven RT: room temperature

7.1.2.6 Characterization of Polymorphic Forms

A total of 5 crystalline forms of Compound 1 were found during thispolymorph screen study. The stack plot of XRPD patterns for these formsare shown in FIG. 1, and the physical characteristics are summarized inTable 8.

TABLE 8 Summary of Characterization Data for Compound 1 PolymorphsRepresentative DSC peak TGA loss DVS or other Form Descriptionconditions (° C.) (wt %) comments A anhydrate intermediate form in 229(onset,  0 ~1.2 wt % water slurries ΔH_(f) = 118 J/g) uptake up to 90%RH: KF: <0.1 wt % B anhydrate Anti-solvent 219 (onset), 224 (exo  0.4(170~230° C.) ~1.4 wt % water recrystallizations peak), 231 (peak)uptake up to 90% RH; KF: <0.1 wt % C anhydrate EQ at 50° C. in 232(onset,  0 ~0.6 wt % water MeCN/water, ΔH_(f) = 126 J/g) uptake up to90% RH; acetone, or KF: <0.1 wt % EtOH/water D solvate recrystallizationin n/a 14.1 14.3% DMSO DMSO/MIBK; (by gas slurry in chromatography)DMSO/anisole E solvate slurry in n/a 19.4 (up to 120° C.), n/aDMSO/MIBK, 24.9 (120~220° C.) DMSO/Anisole or DMSO/IPA n/a: notavailable.

Form A

The XRPD pattern, TGA, DSC, DVS, ¹H NMR, stability profile uponcompression and SEM of Form A of Compound 1 are shown in FIGS. 2-8.

FIG. 2 provides an XRPD pattern of Form A of Compound 1. A list of X-RayDiffraction Peaks for Form A of Compound 1 is provided below in Table 9.

TABLE 9 X-Ray Diffraction Peaks for Form A of Compound 1 Pos. d-spacingRel. Int. No. [°2Th.] [Å] [%] 1 7.23 12.2187 17.6 2 11.52 7.6789 29.7 315.22 5.8209 7.5 4 15.62 5.6720 31.2 5 16.58 5.3466 40.3 6 17.19 5.1576100.0 7 18.08 4.9056 22.3 8 19.00 4.6702 19.6 9 19.60 4.5302 22.1 1021.05 4.2197 29.2 11 21.74 4.0884 8.3 12 22.01 4.0388 7.1 13 22.473.9576 6.0 14 23.22 3.8312 28.6 15 24.17 3.6825 5.6 16 24.77 3.5945 57.217 25.59 3.4813 14.6 18 25.94 3.4356 10.5 19 26.63 3.3470 17.4 20 27.733.2172 10.0 21 28.51 3.1307 7.1 22 29.88 2.9906 19.3 23 30.76 2.9065 7.124 31.59 2.8327 11.1 25 34.82 2.5766 4.8 26 36.05 2.4913 4.3

No TGA weight loss was observed for Form A as depicted in FIG. 3.

A DSC plot for Form A showed a melting event with an onset temperatureof 229° C. and heat of fusion of 118 J/g as depicted in FIG. 4.

A dynamic vapor sorption (DVS) isotherm plot for Form A is shown in FIG.5. When the relative humidity (“RH”) was increased from about 0% toabout 90% RH, Form A exhibited about 1.2% w/w water uptake. Form Acontained less than 0.1% water as determined in a coulometric KarlFischer (KF) titrator equipped with an oven sample processor set at 225°C.

No significant degradation or residual solvent for Form A was observedby ¹H NMR as depicted in FIG. 6.

XRPD pattern for Form A remained substantially unchanged with broaderdiffraction peaks, upon application of 2000-psi pressure for about 1minute as shown in FIG. 7. The SEM picture of Form A is shown in FIG. 8.

Form B

Form B was generated by various anti-solvent recrystallizationexperiments, including MeOH/water, DMSO/IPA, DMSO/toluene, andDMSO/water. Cooling recrystallization from THF/water 1:1 also affordedForm B. The XRPD pattern, SEM, TGA, DSC, DVS, ¹H NMR, and stabilityprofile upon compression of Form B of Compound 1 are shown in FIGS.9-15.

FIG. 9 provides an XRPD pattern of Form B of Compound 1. A list of X-RayDiffraction Peaks for Form B of Compound 1 is provided below in Table10.

TABLE 10 X-Ray Diffraction Peaks for Form B of Compound 1 Pos. d-spacingRel. Int. No. [°2Th.] [Å] [%] 1 7.01 12.6035 9.3 2 11.58 7.6444 8.3 311.80 7.5027 6.8 4 12.73 6.9551 18.4 5 15.38 5.7601 34.8 6 16.32 5.433031.4 7 16.72 5.3012 100.0 8 17.72 5.0046 26.6 9 18.13 4.8930 19.8 1018.77 4.7271 7.5 11 20.41 4.3516 22.0 12 21.02 4.2258 15.9 13 21.214.1881 13.5 14 21.93 4.0529 3.4 15 23.68 3.7581 14.2 16 25.01 3.560110.4 17 25.63 3.4755 37.3 18 26.19 3.4030 9.8 19 26.73 3.3349 8.5 2027.45 3.2499 20.9 21 27.71 3.2193 9.4 22 28.22 3.1623 11.8 23 29.483.0296 4.7 24 30.10 2.9692 15.0 25 31.08 2.8775 18.3 26 31.65 2.8272 6.227 34.29 2.6150 3.4

The SEM picture of Form B is shown in FIG. 10.

No TGA weight loss below 170° C. was observed for Form B. A TGA weightloss of 0.4% was observed between 170˜230° C. as depicted in FIG. 11.

A DSC plot for Form B showed a melt/recrystallization event at 219˜224°C. and a major melting event with a peak temperature of 231° C. asdepicted in FIG. 12.

A dynamic vapor sorption (DVS) isotherm plot for Form B is shown in FIG.13. When the relative humidity (“RH”) was increased from about 0% toabout 90% RH, Form B exhibited about 1.4% w/w water uptake. Form Bcontained less than 0.1% water as determined in a coulometric KarlFischer (KF) titrator equipped with an oven sample processor set at 225°C.

No significant degradation or residual solvent for Form B was observedby ¹H NMR as depicted in FIG. 14.

XRPD pattern for Form B remained substantially unchanged with broaderdiffraction peaks, upon application of 2000-psi pressure for about 1minute as shown in FIG. 15.

Form C

Form C was generated by slurrying in solvent systems containing one ormore of the following solvents: acetonitrile/water, acetone, orethanol/water for extended period of time. Form B, (1×wt) was stirred inacetone (30×vol) at 70-75° C. under nitrogen pressure of 50-55-psi. Thebatch was agitated for at least 24 h. The mixture was cooled to roomtemperature over at least 6 hours. At the end of this period, the batchwas filtered. The cake was washed with acetone (2.3×vol) and dried undervacuum. The solids were co-milled. This afforded a white to off-whitesolid, which is consistent with Form C with expected yield of 85 to 90%and expected LC purity of no less than 98.5% area. The XRPD pattern,SEM, TGA, DSC, DVS, ¹H NMR, and stability profile upon compression ofForm C of Compound 1 are shown in FIGS. 16-22.

FIG. 16 provides an XRPD pattern of Form C of Compound 1. A list ofX-Ray Diffraction Peaks for Form C of Compound 1 is provided below inTable 11.

TABLE 11 X-Ray Diffraction Peaks for Form C of Compound 1 Pos. d-spacingRel. Int. No. [°2Th.] [Å] [%] 1 7.36 12.0091 32.0 2 9.14 9.6750 8.3 311.51 7.6855 44.7 4 12.22 7.2420 4.9 5 15.17 5.8398 8.4 6 15.82 5.601131.8 7 16.68 5.3140 57.1 8 16.92 5.2392 86.8 9 17.72 5.0057 100.0 1018.39 4.8242 21.9 11 19.18 4.6268 36.4 12 19.45 4.5649 27.1 13 21.114.2077 40.4 14 21.82 4.0724 12.4 15 22.28 3.9902 12.0 16 22.57 3.939817.6 17 23.36 3.8082 24.7 18 24.26 3.6695 7.1 19 24.71 3.6026 72.5 2025.74 3.4615 16.9 21 26.03 3.4231 9.7 22 26.51 3.3627 17.7 23 27.883.1998 18.0 24 28.70 3.1104 6.9 25 29.91 2.9871 30.5 26 30.43 2.937510.7 27 30.83 2.9006 5.8 28 32.01 2.7960 16.6 29 37.94 2.3718 5.5

The SEM picture of Form C is shown in FIG. 17.

No TGA weight loss was observed for Form C.

A DSC plot for Form C showed a melting event with an onset temperatureof 232° C. and heat of fusion of 126 J/g as depicted in FIG. 18.

A dynamic vapor sorption (DVS) isotherm plot for Form C is shown in FIG.20. When the relative humidity (“RH”) was increased from about 0% toabout 90% RH, Form C exhibited about 0.6% w/w water uptake. Form Ccontained less than 0.1% water as determined in a coulometric KarlFischer (KF) titrator equipped with an oven sample processor set at 225°C.

No significant degradation or residual solvent for Form C was observedby ¹H NMR as depicted in FIG. 21.

XRPD pattern for Form C remained substantially unchanged with broaderdiffraction peaks, upon application of 2000-psi pressure for about 1minute as shown in FIG. 22.

Form D

Form D of Compound 1 is a DMSO solvate.

Form D was obtained by 1) adding about 1.0 g of Form B to a 100-mLcrystallizer, 2) heating Form B to about 70° C., 3) adding about 10 mLof DMSO/methyl isobutyl ketone (MIBK) (1:2, v/v), 4) adding about 3.8 mLof MIBK to solution over about 20 min, 5) adding about 100 mg of Form Cto the solution, and stirring at about 400 rpm for one day, 6) coolingthe batch to about 20° C. for about 5 h, 0.17° C./min, 7) stirring atabout 20° C. over night, 8) filtering and washing wet cake with about 2mL MIBK, and drying under vacuum at about 45° C.

The XRPD pattern and TGA of Form D of Compound 1 are shown in FIGS.23-24, respectively.

FIG. 23 provides an XRPD pattern of Form D of Compound 1. A list ofX-Ray Diffraction Peaks for Form D of Compound 1 is provided below inTable 12.

TABLE 12 X-Ray Diffraction Peaks for Form D of Compound 1 Pos. d-spacingRel. Int. No. [°2Th.] [Å] [%] 1 4.77 18.5435 3.0 2 9.57 9.2399 7.0 310.55 8.3876 3.1 4 11.95 7.4070 3.7 5 12.50 7.0808 3.5 6 14.06 6.2990100.0 7 14.30 6.1927 92.9 8 16.13 5.4943 3.8 9 17.02 5.2097 8.4 10 17.505.0676 19.8 11 17.78 4.9881 8.0 12 18.09 4.9049 7.7 13 18.27 4.8561 9.014 18.75 4.7326 58.5 15 19.09 4.6482 63.5 16 21.04 4.2228 7.3 17 22.773.9053 10.9 18 23.58 3.7738 53.6 19 24.02 3.7045 24.6 20 24.90 3.57568.4 21 25.22 3.5310 10.0 22 26.37 3.3796 9.4 23 26.63 3.3470 7.9 2428.21 3.1640 5.8 25 29.82 2.9958 3.0 26 30.16 2.9629 5.0 27 30.45 2.93616.7 28 32.48 2.7566 3.3 29 33.03 2.7120 8.1 30 33.69 2.6604 3.4 31 35.322.5413 3.0 32 37.96 2.3702 3.2 33 38.70 2.3269 3.0

A TGA weight loss of about 14.1% up to 140° C. was observed for Form Das depicted in FIG. 24.

Form E

Form E of Compound 1 is a DMSO solvate.

Form E was obtained by 1) adding 200 mg of Form C in to a 3-mL glassvial, 2) adding 0.5 mL of DMSO/MIBK (1:2, v/v) or DMSO/IPA (2:1, v/v)solution, 3) optionally adding Form E seed, 4) stirring at roomtemperature for one day, 5) filtering and washing wet cake with 2 mLMIBK or isopropanol, respectively.

The XRPD pattern and TGA of Form E of Compound 1 are shown in FIGS.25-26, respectively.

FIG. 25 provides an XRPD pattern of Form E of Compound 1. A list ofX-Ray Diffraction Peaks for Form E of Compound 1 is provided below inTable 13.

TABLE 13 X-Ray Diffraction Peaks for Form E of Compound 1 Pos. d-spacingRel. Int. No. [°2Th.] [Å] [%] 1 4.20 21.0329 9.6 2 10.48 8.4394 32.0 312.54 7.0591 28.4 4 14.52 6.1023 9.9 5 15.51 5.7131 17.7 6 16.08 5.5121100.0 7 16.97 5.2256 94.5 8 17.77 4.9908 17.1 9 18.48 4.8001 20.5 1019.54 4.5422 14.7 11 21.15 4.2007 62.8 12 21.72 4.0924 20.8 13 22.643.9270 57.4 14 22.91 3.8826 59.9 15 23.43 3.7977 23.6 16 23.83 3.734823.2 17 24.13 3.6881 29.5 18 25.14 3.5421 35.2 19 26.72 3.3362 49.5 2027.68 3.2232 14.6 21 27.93 3.1949 15.3 22 28.86 3.0942 15.6 23 29.083.0703 18.3 24 30.12 2.9671 7.1 25 30.92 2.8923 12.8 26 32.35 2.7672 5.027 33.21 2.6979 6.9

A TGA weight loss of about 19.4% up to 120° C. was observed for Form Ewith an additional weight loss of 24.9% between 120 and 220° C. asdepicted in FIG. 26.

Amorphous Form

An Amorphous Form of Compound 1 was obtained by 1) adding Compound 1 inTHF and water to form a suspension; 2) heating the suspension to 35-45°C. to obtain a clear solution; 3) freezing the solution to in a coldbath of −78° C.; 4) applying high vacuum to remove the solvent, toobtain an amorphous form of Compound 1.

The DSC thermogram, XRPD pattern, and ¹H NMR spectrum for Amorphous Formof Compound 1 are shown in FIGS. 27-29, respectively.

7.1.3 KG-1 Cell Proliferation Assay

The following is an example of an assay that can be used to determinethe anti-proliferative activity of the solid forms of Compound 1 in KG-1cell line (American Type Culture Collection [ATCC]: catalogue numberATCC® CCL-246™) at 72 hours post-treatment. The seeding density for KG-1can be optimized to ensure assay linearity in 384-well plates.

Increasing concentrations of the solid forms of Compound 1 (0.5 nM to 10μM) are spotted in a 10-point serial dilution fashion (3-fold dilution)in duplicate via an acoustic dispenser (EDC ATS-100) into an empty384-well plate. The dimethyl sulfoxide (DMSO) concentration is keptconstant for a final assay concentration of 0.1% DMSO. Prior to testing,KG-1 cells are grown in RPMI-1640 (Roswell Park Memorial Institute-1640)medium with 10% FBS (fetal bovine serum: HyClone) and expanded inculture flasks to provide sufficient amounts of starting material. Cellsare then diluted to 5000 cells per well, in a 50 μL volume and addeddirectly to the compound-spotted 384-well plates. Cells are allowed togrow for 72 hours in 5% CO₂ at 37° C. At the time when exposure of cellsto compound began (to), initial viable cell number is assessed via CellTiter-Gb® Luminescent Cell Viability Assay at a 1 vol: 2 vol ratioaccording to manufacturer's instructions (Promega Corporation, Madison,Wis.) by quantifying the level of luminescence generated byadenosine-5′-triphosphate (ATP) present in viable cells. After 72 hours,cell viability of the treated cells is assessed via Cell Titer-Gb® andread for luminescence.

While the disclosure has been described with respect to the particularembodiments, it will be apparent to those skilled in the art thatvarious changes and modifications may be made without departing from thespirit and scope of the disclosure as defined in the claims. Suchmodifications are also intended to fall within the scope of the appendedclaims.

All of the patents, patent applications and publications referred toherein are incorporated herein in their entireties. Citation oridentification of any reference in this application is not an admissionthat such reference is available as prior art to this disclosure. Thefull scope of the disclosure is better understood with reference to theappended claims.

What is claimed is:
 1. A solid form of2-(4-chlorophenyl)-N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-2,2-difluoroacetamideor a tautomer thereof:

wherein the solid form is Form A having an X-ray powder diffractionpattern comprising peaks at about 15.6, 16.6, 17.2 or 24.8 degrees 2θ.2. The solid form of claim 1, which is anhydrous.
 3. The solid form ofclaim 1, which is Form A having an X-ray powder diffraction patterncomprising peaks at about 15.6, 16.6, 17.2 and 24.8 degrees 2θ.
 4. Thesolid form of claim 1, having an X-ray powder diffraction patterncomprising peaks at about 11.5, 15.6, 16.6, 17.2, 18.1, 19.0, 19.6,21.1, 23.2 or 24.8 degrees 2θ.
 5. The solid form of claim 3, having anX-ray powder diffraction pattern substantially as shown in FIG.
 2. 6.The solid form of claim 3, having a thermal gravimetric analysis plotsubstantially as shown in FIG.
 3. 7. The solid form of claim 3, having adifferential scanning calorimetry plot comprising an endothermic eventwith an onset temperature of about 229° C.
 8. The solid form of claim 3,having a differential scanning calorimetry plot substantially as shownin FIG.
 4. 9. The solid form of claim 3, which exhibits a mass increaseof less than about 1.2% when subjected to an increase in relativehumidity from about 0% to about 90% relative humidity.
 10. The crystalform of claim 3, which is substantially pure.